WO2024158296A1 - A method for freeze-drying biological matter - Google Patents
A method for freeze-drying biological matter Download PDFInfo
- Publication number
- WO2024158296A1 WO2024158296A1 PCT/NO2024/050018 NO2024050018W WO2024158296A1 WO 2024158296 A1 WO2024158296 A1 WO 2024158296A1 NO 2024050018 W NO2024050018 W NO 2024050018W WO 2024158296 A1 WO2024158296 A1 WO 2024158296A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- flexible receptacle
- receptacle
- biological matter
- freeze
- flexible
- Prior art date
Links
- 238000000034 method Methods 0.000 title claims abstract description 57
- 238000004108 freeze drying Methods 0.000 title claims abstract description 34
- 239000012530 fluid Substances 0.000 claims abstract description 59
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 38
- 238000007789 sealing Methods 0.000 claims abstract description 26
- 238000007710 freezing Methods 0.000 claims abstract description 16
- 230000008014 freezing Effects 0.000 claims abstract description 16
- 238000005520 cutting process Methods 0.000 claims abstract description 13
- 239000007787 solid Substances 0.000 claims abstract description 6
- 210000002381 plasma Anatomy 0.000 claims description 35
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 24
- 239000001301 oxygen Substances 0.000 claims description 24
- 229910052760 oxygen Inorganic materials 0.000 claims description 24
- 239000011888 foil Substances 0.000 claims description 7
- 239000002184 metal Substances 0.000 claims description 7
- 229910052751 metal Inorganic materials 0.000 claims description 7
- 239000007789 gas Substances 0.000 claims description 5
- 239000002245 particle Substances 0.000 claims description 5
- 239000011261 inert gas Substances 0.000 claims description 3
- 230000001590 oxidative effect Effects 0.000 claims description 3
- 239000000463 material Substances 0.000 description 8
- 230000008569 process Effects 0.000 description 8
- 238000009792 diffusion process Methods 0.000 description 6
- 241001465754 Metazoa Species 0.000 description 5
- 239000010836 blood and blood product Substances 0.000 description 4
- 229940125691 blood product Drugs 0.000 description 4
- 238000011109 contamination Methods 0.000 description 4
- 238000003780 insertion Methods 0.000 description 4
- 230000037431 insertion Effects 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 3
- 210000004369 blood Anatomy 0.000 description 3
- 239000008280 blood Substances 0.000 description 3
- 239000012528 membrane Substances 0.000 description 3
- UQSXHKLRYXJYBZ-UHFFFAOYSA-N Iron oxide Chemical compound [Fe]=O UQSXHKLRYXJYBZ-UHFFFAOYSA-N 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 239000012620 biological material Substances 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 239000011521 glass Substances 0.000 description 2
- 230000003647 oxidation Effects 0.000 description 2
- 238000007254 oxidation reaction Methods 0.000 description 2
- -1 polyethylene Polymers 0.000 description 2
- 229920000642 polymer Polymers 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- 102000015081 Blood Coagulation Factors Human genes 0.000 description 1
- 108010039209 Blood Coagulation Factors Proteins 0.000 description 1
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 1
- 239000004277 Ferrous carbonate Substances 0.000 description 1
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 1
- 239000004698 Polyethylene Substances 0.000 description 1
- 239000004743 Polypropylene Substances 0.000 description 1
- 230000002528 anti-freeze Effects 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 239000012472 biological sample Substances 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000003114 blood coagulation factor Substances 0.000 description 1
- 229960004424 carbon dioxide Drugs 0.000 description 1
- 235000011089 carbon dioxide Nutrition 0.000 description 1
- OSQPUMRCKZAIOZ-UHFFFAOYSA-N carbon dioxide;ethanol Chemical compound CCO.O=C=O OSQPUMRCKZAIOZ-UHFFFAOYSA-N 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 238000004891 communication Methods 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 230000008878 coupling Effects 0.000 description 1
- 238000010168 coupling process Methods 0.000 description 1
- 238000005859 coupling reaction Methods 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- RAQDACVRFCEPDA-UHFFFAOYSA-L ferrous carbonate Chemical compound [Fe+2].[O-]C([O-])=O RAQDACVRFCEPDA-UHFFFAOYSA-L 0.000 description 1
- 229960004652 ferrous carbonate Drugs 0.000 description 1
- 235000019268 ferrous carbonate Nutrition 0.000 description 1
- 230000005484 gravity Effects 0.000 description 1
- 230000000887 hydrating effect Effects 0.000 description 1
- 230000036571 hydration Effects 0.000 description 1
- 238000006703 hydration reaction Methods 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 229910000015 iron(II) carbonate Inorganic materials 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 239000004033 plastic Substances 0.000 description 1
- 229920003023 plastic Polymers 0.000 description 1
- 229920000573 polyethylene Polymers 0.000 description 1
- 229920001155 polypropylene Polymers 0.000 description 1
- 229920000915 polyvinyl chloride Polymers 0.000 description 1
- 239000004800 polyvinyl chloride Substances 0.000 description 1
- 238000012414 sterilization procedure Methods 0.000 description 1
- 238000000859 sublimation Methods 0.000 description 1
- 230000008022 sublimation Effects 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N1/00—Preservation of bodies of humans or animals, or parts thereof
- A01N1/10—Preservation of living parts
- A01N1/16—Physical preservation processes
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N1/00—Preservation of bodies of humans or animals, or parts thereof
- A01N1/10—Preservation of living parts
- A01N1/14—Mechanical aspects of preservation; Apparatus or containers therefor
- A01N1/146—Non-refrigerated containers specially adapted for transporting or storing living parts whilst preserving
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61J—CONTAINERS SPECIALLY ADAPTED FOR MEDICAL OR PHARMACEUTICAL PURPOSES; DEVICES OR METHODS SPECIALLY ADAPTED FOR BRINGING PHARMACEUTICAL PRODUCTS INTO PARTICULAR PHYSICAL OR ADMINISTERING FORMS; DEVICES FOR ADMINISTERING FOOD OR MEDICINES ORALLY; BABY COMFORTERS; DEVICES FOR RECEIVING SPITTLE
- A61J1/00—Containers specially adapted for medical or pharmaceutical purposes
- A61J1/05—Containers specially adapted for medical or pharmaceutical purposes for collecting, storing or administering blood, plasma or medical fluids ; Infusion or perfusion containers
- A61J1/10—Bag-type containers
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61J—CONTAINERS SPECIALLY ADAPTED FOR MEDICAL OR PHARMACEUTICAL PURPOSES; DEVICES OR METHODS SPECIALLY ADAPTED FOR BRINGING PHARMACEUTICAL PRODUCTS INTO PARTICULAR PHYSICAL OR ADMINISTERING FORMS; DEVICES FOR ADMINISTERING FOOD OR MEDICINES ORALLY; BABY COMFORTERS; DEVICES FOR RECEIVING SPITTLE
- A61J1/00—Containers specially adapted for medical or pharmaceutical purposes
- A61J1/05—Containers specially adapted for medical or pharmaceutical purposes for collecting, storing or administering blood, plasma or medical fluids ; Infusion or perfusion containers
- A61J1/10—Bag-type containers
- A61J1/12—Bag-type containers with means for holding samples of contents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61J—CONTAINERS SPECIALLY ADAPTED FOR MEDICAL OR PHARMACEUTICAL PURPOSES; DEVICES OR METHODS SPECIALLY ADAPTED FOR BRINGING PHARMACEUTICAL PRODUCTS INTO PARTICULAR PHYSICAL OR ADMINISTERING FORMS; DEVICES FOR ADMINISTERING FOOD OR MEDICINES ORALLY; BABY COMFORTERS; DEVICES FOR RECEIVING SPITTLE
- A61J1/00—Containers specially adapted for medical or pharmaceutical purposes
- A61J1/14—Details; Accessories therefor
- A61J1/20—Arrangements for transferring or mixing fluids, e.g. from vial to syringe
- A61J1/2003—Accessories used in combination with means for transfer or mixing of fluids, e.g. for activating fluid flow, separating fluids, filtering fluid or venting
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/19—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles lyophilised, i.e. freeze-dried, solutions or dispersions
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
- A61M1/00—Suction or pumping devices for medical purposes; Devices for carrying-off, for treatment of, or for carrying-over, body-liquids; Drainage systems
- A61M1/02—Blood transfusion apparatus
- A61M1/0272—Apparatus for treatment of blood or blood constituents prior to or for conservation, e.g. freezing, drying or centrifuging
-
- F—MECHANICAL ENGINEERING; LIGHTING; HEATING; WEAPONS; BLASTING
- F26—DRYING
- F26B—DRYING SOLID MATERIALS OR OBJECTS BY REMOVING LIQUID THEREFROM
- F26B25/00—Details of general application not covered by group F26B21/00 or F26B23/00
- F26B25/06—Chambers, containers, or receptacles
- F26B25/063—Movable containers or receptacles, e.g. carts, trolleys, pallet-boxes
-
- F—MECHANICAL ENGINEERING; LIGHTING; HEATING; WEAPONS; BLASTING
- F26—DRYING
- F26B—DRYING SOLID MATERIALS OR OBJECTS BY REMOVING LIQUID THEREFROM
- F26B5/00—Drying solid materials or objects by processes not involving the application of heat
- F26B5/04—Drying solid materials or objects by processes not involving the application of heat by evaporation or sublimation of moisture under reduced pressure, e.g. in a vacuum
- F26B5/044—Drying solid materials or objects by processes not involving the application of heat by evaporation or sublimation of moisture under reduced pressure, e.g. in a vacuum for drying materials in a batch operation in an enclosure having a plurality of shelves which may be heated
-
- F—MECHANICAL ENGINEERING; LIGHTING; HEATING; WEAPONS; BLASTING
- F26—DRYING
- F26B—DRYING SOLID MATERIALS OR OBJECTS BY REMOVING LIQUID THEREFROM
- F26B5/00—Drying solid materials or objects by processes not involving the application of heat
- F26B5/04—Drying solid materials or objects by processes not involving the application of heat by evaporation or sublimation of moisture under reduced pressure, e.g. in a vacuum
- F26B5/06—Drying solid materials or objects by processes not involving the application of heat by evaporation or sublimation of moisture under reduced pressure, e.g. in a vacuum the process involving freezing
Definitions
- Freeze-drying of various blood products for use in treatment of injured patients has been known since World War II. Freeze-drying of for example blood plasma for later use is thus generally a known process.
- a common way of freeze-drying blood plasma is to freeze dry the blood plasma directly in glass bottles, before sealing the glass bottle with a cap.
- the blood plasma is commonly rehydrated in the bottle, transferred to a separate bag, and injected into for example a patient from the bag.
- the use of multiple receptacles for the blood plasma involves a risk of contamination of the blood plasma during the transfer process, and also a risk of loosing track of the origin of the blood plasma.
- a first aspect of the present invention provides a method for freeze-drying biological matter, the method comprising the steps of introducing a fluid biological matter into a flexible receptacle, closing the flexible receptacle, freezing the fluid biological matter, thereby obtaining a solid frozen biological matter, placing the flexible receptacle in a freeze dryer, lowering the pressure in the freeze dryer to a pressure lower than the vapour pressure of water, cutting an opening in the flexible receptacle such that water vapour may escape the flexible receptacle, freeze drying the frozen biological matter in the freeze dryer, thereby obtaining a freeze-dried biological matter with a predetermined humidity, sealing the flexible receptacle, and removing the flexible receptacle from the freeze dryer.
- the flexible receptacle comprises at least one valve, where the at least one valve is configured to be employed for one or more of introducing the fluid biological matter into the flexible receptacle, introducing water into the flexible receptacle, and removing hydrated freeze-dried biological matter from the flexible receptacle.
- the flexible receptacle comprises a drain valve for removing hydrated freeze-dried biological matter from the flexible receptacle, and a second inlet valve for introducing water into the flexible receptacle.
- the flexible receptacle further comprises a filter arranged adjacent to the drain valve, wherein the filter is configured to block non-hydrated particles of freeze-dried biological matter from flowing through the drain valve.
- the flexible receptacle comprises means for enabling the flexible receptacle to be hung onto a hook, wherein the drain valve is arranged such that the drain valve faces downwards when the flexible receptacle is hung onto a hook.
- the fluid biological matter is introduced into a defined portion of the flexible receptacle in the step of introducing a fluid biological matter into the flexible receptacle.
- the method further comprises the step of delimiting the defined portion of the flexible receptacle by providing one or more temporary seals in the flexible receptacle, preferably by clamping the flexible receptacle.
- the method further comprises a step of introducing an inert gas into the flexible receptacle, and/or introducing a first oxygen scrubber into the flexible receptacle prior to the step of sealing the flexible receptacle.
- the method further comprises the step of mechanically separating two layers of the flexible receptacle after the step of cutting an opening in the flexible receptacle.
- the method further comprises an initial step of positioning the flexible receptacle in a pre-determined position on a carrier.
- the method further comprises the steps of placing the flexible receptacle comprising the freeze-dried biological matter inside an additional receptacle, and sealing the additional receptacle, wherein the additional receptacle optionally comprises a metal foil.
- the method further comprises any one or more of the steps of introducing a second oxygen scrubber into the additional receptacle, evacuating the additional receptacle, and introducing a non-oxidizing gas into the additional receptacle, prior to the step of sealing the additional receptacle.
- the fluid biological matter is human blood plasma.
- the step of sealing the flexible receptacle is performed at a pressure lower than the vapour pressure of water.
- the method further comprises the step of rehydrating the freeze-dried biological matter.
- Figure 1 is a schematic illustration of a flexible receptacle that may be employed in the method according to the present invention
- Figure 2 is a schematic illustration of a plurality of flexible receptacles arranged on a carrier
- Figure 3 is a schematic illustration of a section of a flexible receptacle comprising a hole provided between to seals, where the hole may be employed to hang the flexible receptacle onto a hook,
- Figure 4 is a schematic illustration of a flexible receptacle where the flexible receptacle is arranged within an additional receptacle
- Figure 5 is a schematic illustration of the method for freeze-drying biological matter.
- the present invention provides, as schematically illustrated in figure 5, a method for freeze-drying biological matter.
- the biological matter may here for example be fluid blood plasma or more generally a fluid blood product, for example fluid human blood plasma or a fluid human blood product. More specifically, the biological matter may be a quantity of fluid blood plasma or more generally a quantity of a fluid blood product.
- the method according to the present invention generally relates to the employment of a flexible receptacle for freeze drying biological matter.
- the flexible receptacle may be used as a receptacle for the biological matter prior to freeze drying, as a receptacle for the biological matter during the process of freeze drying, and as a receptacle for the biological matter once freeze dried.
- the method comprises in short the steps of introducing a fluid biological matter into a flexible receptacle, freezing the fluid biological matter, placing the flexible receptacle in a freeze dryer, lowering the pressure in the freeze dryer to a pressure lower than the vapour pressure of water, cutting an opening in the flexible receptacle, freeze drying the frozen biological matter, sealing the flexible receptacle while the flexible receptacle is inside the freeze dryer, and removing the flexible receptacle from the freeze dryer.
- the biological matter thus remains inside the flexible receptacle from the step of introducing the fluid biological matter into the flexible receptacle.
- the latter has been found to be beneficial for reducing contamination as compared to employing methods that are dependent on transferring the biological matter between receptacles.
- the employment of one flexible receptacle as described herein may further be beneficial for tracking the origin or the freeze-dried biological matter, as the flexible receptacle may be marked when the fluid biological matter is being introduced.
- the method according to the present invention comprises an initial step of introducing a fluid biological matter into a flexible receptacle followed by a step of closing the flexible receptacle.
- the fluid biological matter may here be introduced into the flexible receptacle via a first inlet valve or may alternatively be introduced through an opening.
- the step of closing the flexible receptacle may thus involve closing the first inlet valve or by sealing the opening, for example through heat sealing.
- the introduction of the fluid biological matter into the flexible receptacle may be performed by any suitable filling equipment of which there are several commercially available.
- suitable filling equipment examples include equipment that comprises any one of a filling hose, a screw-on coupling for being connected to the first inlet valve, and a rubber plug comprising an insertion canal through which an insertion needle may be introduced.
- the first inlet valve may alternatively comprise such a rubber plug, whereas the filling equipment may comprise an insertion needle for being introduced through the insertion canal.
- the fluid biological matter will according to the method of the present invention undergo a step of freezing the fluid biological matter, thereby resulting in a solid frozen biological matter.
- the freezing of the fluid biological matter may here be performed in a separate freezing unit, and the freezing process may for example be conventional freezing or flash freezing.
- the fluid biological matter may be frozen by lowering the closed flexible receptacle into a dry-ice ethanol bath or a dry-ice antifreeze bath, or by introducing the flexible receptacle into a blast freezer or mechanical freezer.
- alternative methods for freezing may be employed.
- the time between the initial step of introducing the fluid biological matter into a flexible receptacle and the step of freezing the fluid biological matter may generally be kept as short as possible. Particularly when the fluid biological matter is blood plasma it is preferable to perform the step of freezing the blood plasma within 8 hours of the time when the blood plasma was separated from the blood.
- the latter implies that the time between the initial step of introducing the fluid biological matter into a flexible receptacle and the step of freezing the fluid biological matter may preferably be less than 8 hours, preferably less than 6 hours, as some time should be allocated to perform the step of introducing a fluid biological matter into a flexible receptacle and the step of closing the flexible receptacle.
- the then solid frozen biological matter inside the flexible substrate may preferable be kept at a temperature of maximum - 18° C, preferably maximum - 30° C.
- a temperature of - 18° C has been found to be particularly preferable when the solid frozen biological matter is frozen blood plasma, as a storage temperature of maximum - 18° C has been found to preserve coagulation factors in the blood plasma.
- a temperature of - 30° C is generally employed by most blood storage facilities.
- the flexible receptacle containing the frozen biological matter may after the freezing step be introduced into a freeze dryer followed by a step of lowering the pressure in the freeze dryer to a pressure lower than the vapour pressure of water.
- the freeze dryer may thus for example comprise a separate chamber for receiving the flexible receptacle containing the frozen biological matter.
- the freeze dryer may further comprise any one or more of a process condenser, cooling system, vacuum system, and a control unit.
- the freeze dryer may here be configured to perform any one or more of a primary drying step and a secondary drying step.
- the freeze dryer may for example be a contacttype freeze dryer.
- the flexible receptacle 100 containing the frozen biological matter may further, once positioned in the freeze drier and at a pressure lower than the vapour pressure of water, be cut open such that water vapour may escape the flexible receptacle.
- the flexible receptacle 100 may for example be cut open by cutting off a section of the flexible receptacle, for example by cutting along a cut line 150.
- several cuts may be made along separate cut lines 150, thus providing a plurality of openings in the flexible receptacle. Cutting may be made in straight cut line or generally in any type of shape.
- the frozen biological matter will then according to the method of the present invention undergo a step of freeze drying in the freeze dryer until a freeze-dried biological matter with a predetermined humidity is obtained.
- a predetermined humidity may here for example be obtained by performing the step of freeze drying for a predetermined period of time under a set of predetermined conditions, e.g., pressure, temperature and time.
- the predetermined humidity may be obtained by routine optimization for the operating conditions of the freeze drier employed, and the properties and amount of the frozen biological matter to be freeze dried.
- the temperature during freeze drying may further be kept below 0 °C, for example in the range - 20 °C to - 60 °C or alternatively in the range - 40 °C to - 60 °C.
- a cut opening in the flexible receptacle is here preferable to for example using a valve or similar for letting the water out of the flexible receptacle 100, as a cut opening enables a larger opening in the flexible receptacle 100 to be made, a larger diffusion rate for sublimated water, and thus a reduced time required for performing the freeze drying.
- the cut opening may consequently be sized to enable a certain freeze-drying rate and further enable a large surface area the frozen biological matter exposed to the low pressure of the freeze drier. More specifically, a large, exposed surface area of the frozen biological matter will increase sublimation of water molecules from the frozen biological matter during the freeze drying, which may thus result in a higher freeze-drying rate as compared to the use of a similarly sized rigid container where a frozen biological matter would have a smaller exposed surface area.
- the cut opening may be obtained by cutting along a cut line 150 that may be linear, and as schematically illustrated in figure 1, may for example cover one entire side of the flexible receptacle 100.
- the step of lowering the pressure in the freeze dryer to a pressure lower than the vapour pressure of water may herein comprise lowering the pressure in the freeze dryer to a pressure lower than the water pressure caused by the presence of the biological matter.
- the pressure in the freeze dryer may in other words be lowered to a pressure that is lower than the pressure inside the flexible receptacle, more specifically, the pressure in the freeze dryer may be lowered to a pressure that is lower than the vapour pressure of water at the surface of the frozen biological material.
- the pressure in the freeze dryer may for example be provided by arranging a condenser in the freeze dryer that is kept at a temperature that causes water molecules to condense and freeze on the condenser. The vapour pressure of water at the surface of the condenser may thus be kept lower than the water pressure of water at the surface of the frozen biological material.
- the method according to the present invention may optionally include a step of mechanically separating two layers of the flexible receptacle after the step of cutting an opening in the flexible receptacle.
- Said layers may here be two sides of the flexible receptacle that for example may be at least in part stuck together as a consequence of cutting the flexible receptacle, and it may thus be preferable to actively separate any layers stuck together. Separating of any two layers may for example be performed by mechanical means mounted in the freeze dryer, or by built-in gloves.
- the flexible receptacle 100 is according to the present invention sealed while the flexible receptacle 100 is maintained inside the freeze dryer.
- the seal may be made along a seal line 160 that for example may be adjacent to a cut line 150.
- the flexible receptacle 100 may here be sealed at the same pressure in the freeze dryer as was used during the step of freeze drying, or more generally with a pressure in the freeze dryer that is lower than the pressure inside the flexible receptacle.
- the flexible receptacle 100 Sealing the flexible receptacle 100 with a pressure in the freeze dryer being lower than the pressure inside the flexible receptacle has been found to reduce the risk of contamination of the freeze-dried biological matter, as said pressure difference will cause a constant flow of water vapour out of the flexible receptacle as long as the flexible receptacle remains open.
- the pressure difference between the freeze dryer and the interior of the flexible receptacle 100 may in other words contribute to reducing any diffusion of foreign particles into the flexible receptacle.
- the step of sealing the flexible receptacle 100 may here be understood as a process of closing the opening of the flexible receptacle 100 securely.
- the step of sealing the flexible receptacle 100 may be understood as a process where a fluid communication between the freeze-dried biological sample and the surroundings is removed so that free flow of a gas no longer may occur through the opening.
- the step of sealing the flexible receptacle 100 may for example be performed by heat sealing or similar, and may be performed by a sealing unit, such as a heatsealing unit, arranged inside the freeze dryer.
- the flexible receptacle 100 may comprise at least one valve.
- the flexible receptacle 100 may for example comprise a first inlet valve 130 configured to being employed for introducing the fluid biological matter 110 into the flexible receptacle 100.
- the first inlet valve 130 may here be configured to be connected to a suitable filling equipment, for example equipment configured to transfer blood plasma between receptacles.
- the flexible receptacle 100 may additionally or alternatively comprise a second inlet valve 140 configured to being employed for introducing a hydrating fluid, such as water, into the flexible receptacle 100.
- the second inlet valve 140 may thus be employed to hydrate the freeze-dried biological matter subsequent to having performed the method according to the present invention.
- the flexible receptacle 100 may additionally or alternatively comprise a drain valve 170 configured to being employed for extracting hydrated freeze-dried biological matter from the flexible receptacle 100 subsequent to having performed the method according to the present invention.
- the flexible receptacle 100 may, as schematically illustrated in figure 1 comprise both a second inlet valve 140 and a drain valve 170, which may enable hydration of the freeze-dried biological matter to occur simultaneously with extraction of hydrated freeze-dried biological matter from the flexible receptacle 100.
- the presence of a second inlet valve 140 and a drain valve 170 is particularly preferable when the freeze-dried biological matter is freeze-dried blood plasma, and it is desirable to inject said freeze-dried blood plasma into a severely injured human or animal. Water may here be introduced through the second inlet valve 140 simultaneously with extracting a water diluted blood plasma from the drain valve 170.
- the method according to the present invention may thus comprises a step of rehydrating the freeze-dried biological matter, for example using water or more specifically, a saline solution.
- the flexible receptacle 100 may, as schematically illustrated in figure 1, comprise a second inlet valve 140 for introducing water into the flexible receptacle, a drain valve 170 for removing hydrated freeze-dried biological matter from the flexible receptacle, and a filter 180 arranged adjacent to the drain valve.
- the filter 180 is here configured to block non-hydrated particles of freeze-dried biological matter from flowing through the drain valve 170.
- the employment of such a filter 180 is preferable in situations where the freeze-dried biological matter is blood plasma, and the freeze-dried blood plasma is needed in a time of emergency to treat a severely injured human or animal.
- Saline water may here for example be used to hydrate the freeze-dried blood plasma, and the severely injured human or animal may thus initial be injected with mainly saline water before more and more of the freeze-dried blood plasma is hydrated and thereafter injected.
- the filter 180 may in the latter case hinder non-hydrated particles of freeze-dried blood plasma to be injected into the severely injured human or animal.
- the flexible receptacle may generally according to the present invention be a bag, more specifically a blood bag, and may additionally or alternatively be made of a polymer, for example polyethylene, polypropylene, polyvinyl chloride. However, as will be appreciated by a person skilled in the art with knowledge of the present invention the flexible receptacle may be made from other suitable materials.
- the flexible receptacle is preferably made from a material or a plurality of materials that enable the flexible receptacle to be heat sealed and/or transparent.
- the flexible receptacle may be made from a laminate, optionally with at least one layer being made from a metal.
- the flexible receptacle may be cleaned and/or sterilized prior to use or may undergo a cleaning and/or sterilization procedure as a part of its production process and may optionally be stored in a sterile environment prior to use.
- the flexible receptacle 100 may comprise means for enabling the flexible receptacle 100 to be hung onto a hook, and a drain valve 170 for removing hydrated freeze-dried biological matter from the flexible receptacle 100.
- the drain valve 170 is preferably arranged such that the drain valve 170 faces downwards when the flexible receptacle is hung onto a hook, which may enable the hydrated freeze-dried biological matter to be extracted from the flexible receptacle by means of gravity.
- the hydrated freeze- dried biological matter may thus be removed without the need for an external pump or the like, which may be preferable in situations where such equipment is not available, or power is not available.
- the means for enabling the flexible receptacle to be hung onto a hook may for example be a sealed off hole 200 in the flexible receptacle 100, an externally mounted loop, or an externally mounted hook.
- the step of introducing the fluid biological matter 110 into the flexible receptacle 100 may, as schematically illustrated in figure 1, be performed by introducing the fluid biological matter 110 into a defined portion 230 of the flexible receptacle 100, for example through the first inlet valve 130.
- the fluid biological matter 110 may be introduced into a subsection of the flexible receptacle such that the fluid biological matter may only contact a limited portion of the flexible receptacle.
- Introducing the fluid biological 100 matter into a defined portion 230 of the flexible receptacle 100 is preferrable in order to obtain pre-defined shape of frozen biological matter, and later freeze-dried biological matter, but is particularly preferable when the flexible receptacle comprises one or more valves.
- Introducing the fluid biological matter 110 into a defined portion of the flexible receptacle 100 may avoid any contact between the fluid biological matter and any one or more of of a drain valve 170 and a second inlet valve 140 of the flexible receptacle 100. Such contact may be preferrable to avoid, as a contact between the fluid biological matter 110 and a valve may affect the function of that valve, for example during a later rehydration of the biological matter when the biological matter has been freeze-dried.
- a defined portion of the flexible receptacle 100 may be obtained by providing one or more temporary seals 120 in the flexible receptacle 100.
- a temporary seal 120 may here be configured such that the fluid biological matter 110 may not pass the temporary seal 120, and the temporary seal 120 or seals may thus form a pocket, i.e., a defined portion 230, inside the flexible receptacle 100.
- a temporary seal 120 may be provided by clamping the flexible receptacle 100, for example by clamps. Clamping the flexible receptacle 100 may here cause two sides of the flexible receptacle 100 to be forced against each other thereby providing a barrier, /.e. z a temporary seal 120 that the fluid biological matter 110 may not pass.
- FIG. 1 schematically illustrates an example of a defined portion 230 of the flexible receptacle 100 delimited by providing two temporary seals 120, in the flexible receptacle 100.
- a defined portion 230 of the flexible receptacle may thus be obtained and the fluid biological matter 100 may be introduced into the defined portion 230.
- a first inlet valve 130 for introducing the fluid biological matter 110 into the flexible receptacle 110 may be positioned such that the fluid biological matter 110 may be introduced into the defined portion 230 of the flexible receptacle 100.
- Introducing the fluid biological matter 110 into a defined portion 230 may be preferable for avoiding contact between the fluid biological matter 110 and any one or more of a second inlet valve 140, a drain valve 170, and an area of the flexible receptacle where a later sealing may be performed.
- the temporary seal or seals 120 may be broken.
- Introducing the fluid biological matter 110 into a delimited portion may further be preferable in order to enable a homogeneous and/or controlled heat transfer with the later frozen biological matter during freeze drying.
- the method according to the present invention may optionally comprise an additional step of introducing an inert gas into the flexible receptacle prior to sealing the flexible receptacle.
- an introduction of an inter gas into the flexible receptacle has been found to reduce lump formation in the freeze-dried biological matter that may otherwise occur in the freeze-dried biological matter as it is removed from the freeze drier and exposed to an atmospheric pressure.
- a first oxygen scrubber 240 may be placed inside the flexible receptacle 100.
- a first oxygen scrubber 240 may, as schematically illustrated in figure 4 generally be placed inside the flexible receptacle 100 at any point during the method according to the present invention before the step of sealing the flexible receptacle 100 but is preferably introduced after the step of freezing the fluid biological matter 110 or after the step of freeze drying the frozen biological matter.
- An oxygen scrubber may generally herein be one or more pieces of material that irreversibly binds to or combines with oxygen.
- An example of an oxygen scrubber that may be employed is iron powder, such as powdered ferrous oxide or ferrous carbonate.
- Use of a first oxygen scrubber may preferably be used in combination with the flexible receptacle 100 comprising a drain valve 170 and a filter 180 arranged adjacent to the drain valve 170.
- the filter 180 may here be configured to filter out at least parts of the first oxygen scrubber 240.
- a first oxygen scrubber 240 may be applied to reduce the oxidation of the freeze-dried biological matter.
- Some examples include limiting any oxygen diffusion through the flexible receptacle 100 and limiting the availability of oxygen that may diffuse through the flexible receptacle 100. The latter may be particularly preferable if the flexible receptacle 100 is made from a polymer or plastic, as some oxygen diffusion through these materials may occur.
- the method according to the present invention may for example comprise the additional steps of placing the flexible receptacle 100 comprising the freeze-dried biological matter inside an additional receptacle 220 and sealing the additional receptacle 220.
- the additional receptacle may here be flexible, and may optionally comprise a metal foil, e.g., comprise a laminate comprising a metal foil.
- a metal foil may here be preferable, as a metal foil may have a low diffusivity of oxygen.
- the additional receptacle may be made from a specific material or comprise a foil made from a specific material, where said specific material has a lower diffusivity to oxygen than the flexible receptacle.
- an additional receptacle 200 may, prior to the step of sealing the additional receptacle 200, further be combined with any one or more of the steps of introducing a second oxygen scrubber 250 into the additional receptacle 220, evacuating the additional receptacle 220, and introducing a non-oxidizing gas into the additional receptacle 220.
- any diffusion of oxygen through the flexible receptacle 100 may be reduced, which consequently further may reduce the oxidation of the freeze-dried biological matter.
- Figure 4 schematically illustrates a flexible receptacle 100 arranged inside an additional receptacle 220 where a second oxygen scrubber 250 is arranged inside the additional receptacle 220.
- the handling and transport of the flexible receptacle 100 may generally throughout the method according to the present invention be manual, or automatic.
- the flexible receptacle 100 may for example be aligned according to a suitable cutting tool and aligned according to a filling unit for introducing a fluid biological matter into the flexible receptable 100.
- the flexible receptable 100 may thus be positioned with a certain arrangement by suitable handling means, such as a robotic handling and transport system, such that further manual interaction with the flexible receptacle 100 may be avoided before the method is completed.
- suitable handling means such as a robotic handling and transport system, such that further manual interaction with the flexible receptacle 100 may be avoided before the method is completed.
- the latter may have the advantage of reducing the risk of contamination of the biological matter.
- the method may further comprise an initial step of positioning the flexible receptacle 100 in a pre-determined position on a carrier 210.
- the carrier 210 may be handled manually or automatically, i.e., be moved in and out of the freeze drier etc. and may further facilitate for the handling of multiple flexible receptacles at once.
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Abstract
The invention describes a method for freeze-drying biological matter (110), the method comprising the steps of introducing a fluid biological matter (110) into a flexible receptacle (100), closing the flexible receptacle (100), freezing the fluid biological matter (110), thereby obtaining a solid frozen biological matter (110), placing the flexible receptacle (100) in a freeze dryer, lowering the pressure in the freeze dryer to a pressure lower than the vapour pressure of water, cutting an opening in the flexible receptacle (100) such that water vapour may escape the flexible receptacle (100), freeze drying the frozen biological matter (110) in the freeze dryer, thereby obtaining a freeze-dried biological matter (110) with a predetermined humidity, sealing the flexible receptacle (100), and removing the flexible receptacle (100) from the freeze dryer
Description
A method for freeze-drying biological matter
Field of invention
The invention relates to the field of freeze-drying of biological matter
Background
[0001] Freeze-drying of various blood products for use in treatment of injured patients has been known since World War II. Freeze-drying of for example blood plasma for later use is thus generally a known process.
[0002] A common way of freeze-drying blood plasma is to freeze dry the blood plasma directly in glass bottles, before sealing the glass bottle with a cap. When the blood plasma is needed, the blood plasma is commonly rehydrated in the bottle, transferred to a separate bag, and injected into for example a patient from the bag. The use of multiple receptacles for the blood plasma involves a risk of contamination of the blood plasma during the transfer process, and also a risk of loosing track of the origin of the blood plasma.
[0003] Another way of freeze-drying blood plasma is described in US20090223080A1. Here, two chambers are employed with a connector connecting the two chambers, with a permeable membrane being positioned in one chamber and with the frozen blood plasma to be freeze dried being positioned in the other chamber prior to freeze-drying. During freeze-drying the connection between the two chambers remain open and water vapour may thus diffuse from one chamber to the other, whereas when the freeze-drying ends the connection is closed. A drawback with this solution, however, is that the limited path for diffusion of water vapour and the limited diffusivity of the membrane makes the freeze-drying process slow. US5309649 A describes a solution where a membrane is located above the blood plasma.
[0004] It is an aim of the present invention to address at least some of the issues with the prior art.
Summary of the invention
[0005] A first aspect of the present invention provides a method for freeze-drying biological matter, the method comprising the steps of introducing a fluid biological matter into a flexible receptacle, closing the flexible receptacle, freezing the fluid biological matter, thereby obtaining a solid frozen biological matter, placing the flexible receptacle in a freeze dryer, lowering the pressure in the freeze dryer to a pressure lower than the vapour pressure of water, cutting an opening in the flexible receptacle such that water vapour may escape the flexible receptacle, freeze drying the frozen biological matter in the freeze dryer,
thereby obtaining a freeze-dried biological matter with a predetermined humidity, sealing the flexible receptacle, and removing the flexible receptacle from the freeze dryer.
[0006] In an embodiment of the invention the flexible receptacle comprises at least one valve, where the at least one valve is configured to be employed for one or more of introducing the fluid biological matter into the flexible receptacle, introducing water into the flexible receptacle, and removing hydrated freeze-dried biological matter from the flexible receptacle.
[0007] In another embodiment of the invention the flexible receptacle comprises a drain valve for removing hydrated freeze-dried biological matter from the flexible receptacle, and a second inlet valve for introducing water into the flexible receptacle.
[0008] In yet another embodiment of the invention the flexible receptacle further comprises a filter arranged adjacent to the drain valve, wherein the filter is configured to block non-hydrated particles of freeze-dried biological matter from flowing through the drain valve.
[0009] In yet another embodiment of the invention the flexible receptacle comprises means for enabling the flexible receptacle to be hung onto a hook, wherein the drain valve is arranged such that the drain valve faces downwards when the flexible receptacle is hung onto a hook.
[0010] In yet another embodiment of the invention the fluid biological matter is introduced into a defined portion of the flexible receptacle in the step of introducing a fluid biological matter into the flexible receptacle.
[0011] In yet another embodiment of the invention the method further comprises the step of delimiting the defined portion of the flexible receptacle by providing one or more temporary seals in the flexible receptacle, preferably by clamping the flexible receptacle.
[0012] In yet another embodiment of the invention the method further comprises a step of introducing an inert gas into the flexible receptacle, and/or introducing a first oxygen scrubber into the flexible receptacle prior to the step of sealing the flexible receptacle.
[0013] In yet another embodiment of the invention the method further comprises the step of mechanically separating two layers of the flexible receptacle after the step of cutting an opening in the flexible receptacle.
[0014] In yet another embodiment of the invention the method further comprises an initial step of positioning the flexible receptacle in a pre-determined position on a carrier.
[0015] In yet another embodiment of the invention the method further comprises the steps of placing the flexible receptacle comprising the freeze-dried biological matter inside an additional receptacle, and sealing the additional receptacle, wherein the additional receptacle optionally comprises a metal foil.
[0016] In yet another embodiment of the invention the method further comprises any one or more of the steps of introducing a second oxygen scrubber into the additional receptacle, evacuating the additional receptacle, and introducing a non-oxidizing gas into the additional receptacle, prior to the step of sealing the additional receptacle.
[0017] In yet another embodiment of the invention the fluid biological matter is human blood plasma.
[0018] In yet another embodiment of the invention the step of sealing the flexible receptacle is performed at a pressure lower than the vapour pressure of water.
[0019] In yet another embodiment of the invention the method further comprises the step of rehydrating the freeze-dried biological matter.
Brief description of the drawings
[0020] Figure 1 is a schematic illustration of a flexible receptacle that may be employed in the method according to the present invention,
[0021] Figure 2 is a schematic illustration of a plurality of flexible receptacles arranged on a carrier,
[0022] Figure 3 is a schematic illustration of a section of a flexible receptacle comprising a hole provided between to seals, where the hole may be employed to hang the flexible receptacle onto a hook,
[0023] Figure 4 is a schematic illustration of a flexible receptacle where the flexible receptacle is arranged within an additional receptacle,
[0024] Figure 5 is a schematic illustration of the method for freeze-drying biological matter.
Detailed description of the invention
[0025] In the following, general embodiments as well as particular exemplary embodiments of the invention will be described. References will be made to the accompanying drawings. It shall be noted, however, that the drawings are exemplary embodiments only, and that other features and embodiments may well be within the scope of the invention as claimed. Further, the mentioning of references such as "a" or "an" etc. should not be construed as excluding a plurality, and the term "disclosure" may herein be used interchangeably with the term "invention".
[0026] Unless otherwise defined, all terms of art, notations and other scientific terms or terminology used herein are intended to have the meanings commonly understood by those of skill in the art to which this invention pertains. Certain terms of art, notations, and other scientific terms or terminology may, however, be defined specifically as indicated below.
[0027] The present invention provides, as schematically illustrated in figure 5, a method for freeze-drying biological matter. The biological matter may here for example be fluid blood plasma or more generally a fluid blood product, for example fluid human blood plasma or a fluid human blood product. More specifically, the biological matter may be a quantity of fluid blood plasma or more generally a quantity of a fluid blood product.
[0028] The method according to the present invention generally relates to the employment of a flexible receptacle for freeze drying biological matter. The flexible receptacle may be used as a receptacle for the biological matter prior to freeze drying, as a receptacle for the biological matter during the process of freeze drying, and as a receptacle for the biological matter once freeze dried. The method comprises in short the steps of introducing a fluid biological matter into a flexible receptacle, freezing the fluid biological matter, placing the flexible receptacle in a freeze dryer, lowering the pressure in the freeze dryer to a pressure lower than the vapour pressure of water, cutting an opening in the flexible receptacle, freeze drying the frozen biological matter, sealing the flexible receptacle while the flexible receptacle is inside the freeze dryer, and removing the flexible receptacle from the freeze dryer. The biological matter thus remains inside the flexible receptacle from the step of introducing the fluid biological matter into the flexible receptacle. The latter has been found to be beneficial for reducing contamination as compared to employing methods that are dependent on transferring the biological matter between receptacles. The employment of one flexible receptacle as described herein may further be beneficial for tracking the origin or the freeze-dried biological matter, as the flexible receptacle may be marked when the fluid biological matter is being introduced.
[0029] In particular, the method according to the present invention comprises an initial step of introducing a fluid biological matter into a flexible receptacle followed by a step of closing the flexible receptacle. The fluid biological matter may here be introduced into the flexible receptacle via a first inlet valve or may alternatively be introduced through an opening. The step of closing the flexible receptacle may thus involve closing the first inlet valve or by sealing the opening, for example through heat sealing. As will be appreciated by a person skilled in the art with knowledge of the present invention, the introduction of the fluid
biological matter into the flexible receptacle may be performed by any suitable filling equipment of which there are several commercially available. Examples of suitable filling equipment include equipment that comprises any one of a filling hose, a screw-on coupling for being connected to the first inlet valve, and a rubber plug comprising an insertion canal through which an insertion needle may be introduced. The first inlet valve may alternatively comprise such a rubber plug, whereas the filling equipment may comprise an insertion needle for being introduced through the insertion canal.
[0030] Once inside the flexible receptacle, the fluid biological matter will according to the method of the present invention undergo a step of freezing the fluid biological matter, thereby resulting in a solid frozen biological matter. The freezing of the fluid biological matter may here be performed in a separate freezing unit, and the freezing process may for example be conventional freezing or flash freezing. As a way of example, the fluid biological matter may be frozen by lowering the closed flexible receptacle into a dry-ice ethanol bath or a dry-ice antifreeze bath, or by introducing the flexible receptacle into a blast freezer or mechanical freezer. A person skilled in the art with knowledge of the present invention will appreciate that alternative methods for freezing may be employed.
[0031] The time between the initial step of introducing the fluid biological matter into a flexible receptacle and the step of freezing the fluid biological matter may generally be kept as short as possible. Particularly when the fluid biological matter is blood plasma it is preferable to perform the step of freezing the blood plasma within 8 hours of the time when the blood plasma was separated from the blood. The latter implies that the time between the initial step of introducing the fluid biological matter into a flexible receptacle and the step of freezing the fluid biological matter may preferably be less than 8 hours, preferably less than 6 hours, as some time should be allocated to perform the step of introducing a fluid biological matter into a flexible receptacle and the step of closing the flexible receptacle.
[0032] Once frozen, before being freeze dried, the then solid frozen biological matter inside the flexible substrate may preferable be kept at a temperature of maximum - 18° C, preferably maximum - 30° C. A temperature of - 18° C has been found to be particularly preferable when the solid frozen biological matter is frozen blood plasma, as a storage temperature of maximum - 18° C has been found to preserve coagulation factors in the blood plasma. A temperature of - 30° C is generally employed by most blood storage facilities.
[0033] The flexible receptacle containing the frozen biological matter may after the freezing step be introduced into a freeze dryer followed by a step of lowering the
pressure in the freeze dryer to a pressure lower than the vapour pressure of water. The freeze dryer may thus for example comprise a separate chamber for receiving the flexible receptacle containing the frozen biological matter. As will be appreciated by a person skilled in the art with knowledge of the present invention the freeze dryer may further comprise any one or more of a process condenser, cooling system, vacuum system, and a control unit. The freeze dryer may here be configured to perform any one or more of a primary drying step and a secondary drying step. The freeze dryer may for example be a contacttype freeze dryer.
[0034] The flexible receptacle 100 containing the frozen biological matter may further, once positioned in the freeze drier and at a pressure lower than the vapour pressure of water, be cut open such that water vapour may escape the flexible receptacle. As schematically illustrated in figure 1, the flexible receptacle 100 may for example be cut open by cutting off a section of the flexible receptacle, for example by cutting along a cut line 150. Alternatively, several cuts may be made along separate cut lines 150, thus providing a plurality of openings in the flexible receptacle. Cutting may be made in straight cut line or generally in any type of shape. Once opened, the frozen biological matter will then according to the method of the present invention undergo a step of freeze drying in the freeze dryer until a freeze-dried biological matter with a predetermined humidity is obtained. A predetermined humidity may here for example be obtained by performing the step of freeze drying for a predetermined period of time under a set of predetermined conditions, e.g., pressure, temperature and time. As will be appreciated by a person skilled in the art, the predetermined humidity may be obtained by routine optimization for the operating conditions of the freeze drier employed, and the properties and amount of the frozen biological matter to be freeze dried. As will further be appreciated, the temperature during freeze drying may further be kept below 0 °C, for example in the range - 20 °C to - 60 °C or alternatively in the range - 40 °C to - 60 °C.
[0035] During the step of freeze drying the frozen biological matter, water molecules will sublimate from the frozen biological matter and then diffuse out of the flexible receptacle 100 through the cut opening, before optionally being evacuated out of the freeze dryer or condensed inside the freeze dryer using a condenser. The use of a cut opening in the flexible receptacle is here preferable to for example using a valve or similar for letting the water out of the flexible receptacle 100, as a cut opening enables a larger opening in the flexible receptacle 100 to be made, a larger diffusion rate for sublimated water, and thus a reduced time required for performing the freeze drying. The cut opening may
consequently be sized to enable a certain freeze-drying rate and further enable a large surface area the frozen biological matter exposed to the low pressure of the freeze drier. More specifically, a large, exposed surface area of the frozen biological matter will increase sublimation of water molecules from the frozen biological matter during the freeze drying, which may thus result in a higher freeze-drying rate as compared to the use of a similarly sized rigid container where a frozen biological matter would have a smaller exposed surface area. The cut opening may be obtained by cutting along a cut line 150 that may be linear, and as schematically illustrated in figure 1, may for example cover one entire side of the flexible receptacle 100.
[0036] The step of lowering the pressure in the freeze dryer to a pressure lower than the vapour pressure of water may herein comprise lowering the pressure in the freeze dryer to a pressure lower than the water pressure caused by the presence of the biological matter. The pressure in the freeze dryer may in other words be lowered to a pressure that is lower than the pressure inside the flexible receptacle, more specifically, the pressure in the freeze dryer may be lowered to a pressure that is lower than the vapour pressure of water at the surface of the frozen biological material. The pressure in the freeze dryer may for example be provided by arranging a condenser in the freeze dryer that is kept at a temperature that causes water molecules to condense and freeze on the condenser. The vapour pressure of water at the surface of the condenser may thus be kept lower than the water pressure of water at the surface of the frozen biological material.
[0037] The method according to the present invention may optionally include a step of mechanically separating two layers of the flexible receptacle after the step of cutting an opening in the flexible receptacle. Said layers may here be two sides of the flexible receptacle that for example may be at least in part stuck together as a consequence of cutting the flexible receptacle, and it may thus be preferable to actively separate any layers stuck together. Separating of any two layers may for example be performed by mechanical means mounted in the freeze dryer, or by built-in gloves.
[0038] Once a freeze-dried biological matter with a predetermined humidity is obtained, the flexible receptacle 100 is according to the present invention sealed while the flexible receptacle 100 is maintained inside the freeze dryer. As schematically illustrated in figure 1, the seal may be made along a seal line 160 that for example may be adjacent to a cut line 150. The flexible receptacle 100 may here be sealed at the same pressure in the freeze dryer as was used during the step of freeze drying, or more generally with a pressure in the freeze dryer that is
lower than the pressure inside the flexible receptacle. Sealing the flexible receptacle 100 with a pressure in the freeze dryer being lower than the pressure inside the flexible receptacle has been found to reduce the risk of contamination of the freeze-dried biological matter, as said pressure difference will cause a constant flow of water vapour out of the flexible receptacle as long as the flexible receptacle remains open. The pressure difference between the freeze dryer and the interior of the flexible receptacle 100 may in other words contribute to reducing any diffusion of foreign particles into the flexible receptacle. As will be appreciated by a person skilled in the art, the step of sealing the flexible receptacle 100 may here be understood as a process of closing the opening of the flexible receptacle 100 securely. More specifically, the step of sealing the flexible receptacle 100 may be understood as a process where a fluid communication between the freeze-dried biological sample and the surroundings is removed so that free flow of a gas no longer may occur through the opening. The step of sealing the flexible receptacle 100 may for example be performed by heat sealing or similar, and may be performed by a sealing unit, such as a heatsealing unit, arranged inside the freeze dryer.
[0039] As schematically illustrated in figure 1 the flexible receptacle 100 may comprise at least one valve. The flexible receptacle 100 may for example comprise a first inlet valve 130 configured to being employed for introducing the fluid biological matter 110 into the flexible receptacle 100. The first inlet valve 130 may here be configured to be connected to a suitable filling equipment, for example equipment configured to transfer blood plasma between receptacles. The flexible receptacle 100 may additionally or alternatively comprise a second inlet valve 140 configured to being employed for introducing a hydrating fluid, such as water, into the flexible receptacle 100. The second inlet valve 140 may thus be employed to hydrate the freeze-dried biological matter subsequent to having performed the method according to the present invention. The flexible receptacle 100 may additionally or alternatively comprise a drain valve 170 configured to being employed for extracting hydrated freeze-dried biological matter from the flexible receptacle 100 subsequent to having performed the method according to the present invention.
[0040] The flexible receptacle 100 may, as schematically illustrated in figure 1 comprise both a second inlet valve 140 and a drain valve 170, which may enable hydration of the freeze-dried biological matter to occur simultaneously with extraction of hydrated freeze-dried biological matter from the flexible receptacle 100. The presence of a second inlet valve 140 and a drain valve 170 is particularly preferable when the freeze-dried biological matter is freeze-dried
blood plasma, and it is desirable to inject said freeze-dried blood plasma into a severely injured human or animal. Water may here be introduced through the second inlet valve 140 simultaneously with extracting a water diluted blood plasma from the drain valve 170. As it will inevitably take some time to dissolve the freeze-dried blood plasma, injection of diluted blood plasma into the severely injured human or animal may start immediately, where the concentration of the diluted blood plasma increases over time until the freeze-dried blood plasma is fully hydrated. The method according to the present invention may thus comprises a step of rehydrating the freeze-dried biological matter, for example using water or more specifically, a saline solution.
[0041] The flexible receptacle 100 may, as schematically illustrated in figure 1, comprise a second inlet valve 140 for introducing water into the flexible receptacle, a drain valve 170 for removing hydrated freeze-dried biological matter from the flexible receptacle, and a filter 180 arranged adjacent to the drain valve. The filter 180 is here configured to block non-hydrated particles of freeze-dried biological matter from flowing through the drain valve 170. The employment of such a filter 180 is preferable in situations where the freeze-dried biological matter is blood plasma, and the freeze-dried blood plasma is needed in a time of emergency to treat a severely injured human or animal. Saline water may here for example be used to hydrate the freeze-dried blood plasma, and the severely injured human or animal may thus initial be injected with mainly saline water before more and more of the freeze-dried blood plasma is hydrated and thereafter injected. The filter 180 may in the latter case hinder non-hydrated particles of freeze-dried blood plasma to be injected into the severely injured human or animal.
[0042] The flexible receptacle may generally according to the present invention be a bag, more specifically a blood bag, and may additionally or alternatively be made of a polymer, for example polyethylene, polypropylene, polyvinyl chloride. However, as will be appreciated by a person skilled in the art with knowledge of the present invention the flexible receptacle may be made from other suitable materials. The flexible receptacle is preferably made from a material or a plurality of materials that enable the flexible receptacle to be heat sealed and/or transparent. The flexible receptacle may be made from a laminate, optionally with at least one layer being made from a metal. The flexible receptacle may be cleaned and/or sterilized prior to use or may undergo a cleaning and/or sterilization procedure as a part of its production process and may optionally be stored in a sterile environment prior to use.
[0043] As schematically illustrated in figure 3, the flexible receptacle 100 may comprise means for enabling the flexible receptacle 100 to be hung onto a hook, and a drain valve 170 for removing hydrated freeze-dried biological matter from the flexible receptacle 100. Here the drain valve 170 is preferably arranged such that the drain valve 170 faces downwards when the flexible receptacle is hung onto a hook, which may enable the hydrated freeze-dried biological matter to be extracted from the flexible receptacle by means of gravity. The hydrated freeze- dried biological matter may thus be removed without the need for an external pump or the like, which may be preferable in situations where such equipment is not available, or power is not available. The means for enabling the flexible receptacle to be hung onto a hook may for example be a sealed off hole 200 in the flexible receptacle 100, an externally mounted loop, or an externally mounted hook.
[0044] The step of introducing the fluid biological matter 110 into the flexible receptacle 100 may, as schematically illustrated in figure 1, be performed by introducing the fluid biological matter 110 into a defined portion 230 of the flexible receptacle 100, for example through the first inlet valve 130. In other words, the fluid biological matter 110 may be introduced into a subsection of the flexible receptacle such that the fluid biological matter may only contact a limited portion of the flexible receptacle. Introducing the fluid biological 100 matter into a defined portion 230 of the flexible receptacle 100 is preferrable in order to obtain pre-defined shape of frozen biological matter, and later freeze-dried biological matter, but is particularly preferable when the flexible receptacle comprises one or more valves. Introducing the fluid biological matter 110 into a defined portion of the flexible receptacle 100 may avoid any contact between the fluid biological matter and any one or more of of a drain valve 170 and a second inlet valve 140 of the flexible receptacle 100. Such contact may be preferrable to avoid, as a contact between the fluid biological matter 110 and a valve may affect the function of that valve, for example during a later rehydration of the biological matter when the biological matter has been freeze-dried.
[0045] As schematically illustrated in figure 1, a defined portion of the flexible receptacle 100 may be obtained by providing one or more temporary seals 120 in the flexible receptacle 100. A temporary seal 120 may here be configured such that the fluid biological matter 110 may not pass the temporary seal 120, and the temporary seal 120 or seals may thus form a pocket, i.e., a defined portion 230, inside the flexible receptacle 100. A temporary seal 120 may be provided by clamping the flexible receptacle 100, for example by clamps. Clamping the flexible receptacle 100 may here cause two sides of the flexible
receptacle 100 to be forced against each other thereby providing a barrier, /.e.z a temporary seal 120 that the fluid biological matter 110 may not pass. The clamping may here be performed by any number of suitable external clamps, for example flat clamps, optionally mounted with a rubber section for contacting the flexible receptacle 100. Figure 1 schematically illustrates an example of a defined portion 230 of the flexible receptacle 100 delimited by providing two temporary seals 120, in the flexible receptacle 100. A defined portion 230 of the flexible receptacle may thus be obtained and the fluid biological matter 100 may be introduced into the defined portion 230. As also illustrated in figure 1, a first inlet valve 130 for introducing the fluid biological matter 110 into the flexible receptacle 110 may be positioned such that the fluid biological matter 110 may be introduced into the defined portion 230 of the flexible receptacle 100. Introducing the fluid biological matter 110 into a defined portion 230 may be preferable for avoiding contact between the fluid biological matter 110 and any one or more of a second inlet valve 140, a drain valve 170, and an area of the flexible receptacle where a later sealing may be performed. Once the fluid biological matter 110 is frozen, the temporary seal or seals 120 may be broken. Introducing the fluid biological matter 110 into a delimited portion may further be preferable in order to enable a homogeneous and/or controlled heat transfer with the later frozen biological matter during freeze drying.
[0046] The method according to the present invention may optionally comprise an additional step of introducing an inert gas into the flexible receptacle prior to sealing the flexible receptacle. Such an introduction of an inter gas into the flexible receptacle has been found to reduce lump formation in the freeze-dried biological matter that may otherwise occur in the freeze-dried biological matter as it is removed from the freeze drier and exposed to an atmospheric pressure.
[0047] Residual water and oxygen are generally considered factors that limit the shelf life of the freeze-dried biological matter, particularly freeze-dried blood plasma. Oxygen may react with the freeze-dried biological matter and may cause the freeze-dried biological matter to deteriorate. In order to limit the availability of oxygen inside a sealed flexible receptacle, a first oxygen scrubber 240 may be placed inside the flexible receptacle 100. A first oxygen scrubber 240 may, as schematically illustrated in figure 4 generally be placed inside the flexible receptacle 100 at any point during the method according to the present invention before the step of sealing the flexible receptacle 100 but is preferably introduced after the step of freezing the fluid biological matter 110 or after the step of freeze drying the frozen biological matter. An oxygen scrubber may generally herein be one or more pieces of material that irreversibly binds to or
combines with oxygen. An example of an oxygen scrubber that may be employed is iron powder, such as powdered ferrous oxide or ferrous carbonate. Use of a first oxygen scrubber may preferably be used in combination with the flexible receptacle 100 comprising a drain valve 170 and a filter 180 arranged adjacent to the drain valve 170. The filter 180 may here be configured to filter out at least parts of the first oxygen scrubber 240.
[0048] Additionally, or alternatively to introducing a first oxygen scrubber 240 inside the flexible receptacle 100, other means may be applied to reduce the oxidation of the freeze-dried biological matter. Some examples include limiting any oxygen diffusion through the flexible receptacle 100 and limiting the availability of oxygen that may diffuse through the flexible receptacle 100. The latter may be particularly preferable if the flexible receptacle 100 is made from a polymer or plastic, as some oxygen diffusion through these materials may occur. The method according to the present invention may for example comprise the additional steps of placing the flexible receptacle 100 comprising the freeze-dried biological matter inside an additional receptacle 220 and sealing the additional receptacle 220. The additional receptacle may here be flexible, and may optionally comprise a metal foil, e.g., comprise a laminate comprising a metal foil. The use of a metal foil may here be preferable, as a metal foil may have a low diffusivity of oxygen. Optionally or alternatively, the additional receptacle may be made from a specific material or comprise a foil made from a specific material, where said specific material has a lower diffusivity to oxygen than the flexible receptacle. The employment of an additional receptacle 200 may, prior to the step of sealing the additional receptacle 200, further be combined with any one or more of the steps of introducing a second oxygen scrubber 250 into the additional receptacle 220, evacuating the additional receptacle 220, and introducing a non-oxidizing gas into the additional receptacle 220. By reducing the amount of oxygen inside the sealed additional receptacle 200, any diffusion of oxygen through the flexible receptacle 100 may be reduced, which consequently further may reduce the oxidation of the freeze-dried biological matter. Figure 4 schematically illustrates a flexible receptacle 100 arranged inside an additional receptacle 220 where a second oxygen scrubber 250 is arranged inside the additional receptacle 220.
[0049] The handling and transport of the flexible receptacle 100 may generally throughout the method according to the present invention be manual, or automatic. The flexible receptacle 100 may for example be aligned according to a suitable cutting tool and aligned according to a filling unit for introducing a fluid biological matter into the flexible receptable 100. The flexible receptable
100 may thus be positioned with a certain arrangement by suitable handling means, such as a robotic handling and transport system, such that further manual interaction with the flexible receptacle 100 may be avoided before the method is completed. The latter may have the advantage of reducing the risk of contamination of the biological matter. As schematically illustrated in figure 2, the method may further comprise an initial step of positioning the flexible receptacle 100 in a pre-determined position on a carrier 210. The carrier 210 may be handled manually or automatically, i.e., be moved in and out of the freeze drier etc. and may further facilitate for the handling of multiple flexible receptacles at once.
Claims
1. A method for freeze-drying biological matter (110), the method comprising the steps of introducing a fluid biological matter (110) into a flexible receptacle (100), closing the flexible receptacle (100), freezing the fluid biological matter (110), thereby obtaining a solid frozen biological matter (110), placing the flexible receptacle (100) in a freeze dryer, lowering the pressure in the freeze dryer to a pressure lower than the vapour pressure of water, cutting an opening in the flexible receptacle (100) such that water vapour may escape the flexible receptacle (100), freeze drying the frozen biological matter (110) in the freeze dryer, thereby obtaining a freeze-dried biological matter (110) with a predetermined humidity, sealing the flexible receptacle (100), and removing the flexible receptacle (100) from the freeze dryer.
2. The method according to claim 1, wherein the flexible receptacle (100) comprises at least one valve, where the at least one valve is configured to be employed for one or more of introducing the fluid biological matter (110) into the flexible receptacle (100), introducing water into the flexible receptacle (100), and removing hydrated freeze-dried biological matter (110) from the flexible receptacle (100).
3. The method according to claim 2, wherein the flexible receptacle (100) comprises a drain valve (170) for removing hydrated freeze-dried biological matter (110) from the flexible receptacle (100), and a second inlet valve (140) for introducing water into the flexible receptacle (100).
4. The method according to claim 3, wherein the flexible receptacle (100) further comprises a filter (180) arranged adjacent to the drain valve (170),
wherein the filter (180) is configured to block non-hydrated particles of freeze- dried biological matter (110) from flowing through the drain valve (170).
5. The method according to any one of the claims 3 or 4, wherein the flexible receptacle (100) comprises means for enabling the flexible receptacle (100) to be hung onto a hook, wherein the drain valve (170) is arranged such that the drain valve (170) faces downwards when the flexible receptacle (100) is hung onto the hook.
6. The method according to any one of the preceding claims, wherein in the step of introducing a fluid biological matter (110) into the flexible receptacle (100), the fluid biological matter (110) is introduced into a defined portion of the flexible receptacle (100).
7. The method according to claim 6, further comprising the step of delimiting the defined portion of the flexible receptacle (100) by providing one or more temporary seals (120) in the flexible receptacle (100), preferably by clamping the flexible receptacle (100).
8. The method according to any one of the preceding claims, further comprising a step of introducing an inert gas into the flexible receptacle (100), and/or introducing a first oxygen scrubber (240) into the flexible receptacle (100) prior to the step of sealing the flexible receptacle (100).
9. The method according to any one of the preceding claims, further comprising the step of mechanically separating two layers of the flexible receptacle (100) after the step of cutting an opening in the flexible receptacle (100).
10. The method according to any one of the preceding claims, further comprising an initial step of positioning the flexible receptacle (100) in a pre-determined position on a carrier (210).
11. The method according to any one of the preceding claims, further comprising the steps of placing the flexible receptacle (100) comprising the freeze-dried biological matter (110) inside an additional receptacle (220), and sealing the additional receptacle (220), wherein the additional receptacle (220) optionally comprises a metal foil.
12. The method according to claim 11, further comprising any one or more of the steps of introducing a second oxygen scrubber (250) into the additional receptacle (220), evacuating the additional receptacle (220), and introducing a non-oxidizing gas into the additional receptacle (220), prior to the step of sealing the additional receptacle (220).
13. The method according to any one of the preceding claims, wherein the fluid biological matter is human blood plasma.
14. The method according to any one of the preceding claims, wherein the step of sealing the flexible receptacle (100) is performed at a pressure lower than the vapour pressure of water.
15. The method according to any one of the preceding claims, further comprising the step of rehydrating the freeze-dried biological matter (110).
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| NO20230074A NO20230074A1 (en) | 2023-01-26 | 2023-01-26 | Method for safe and effective use of freeze-dried material |
| NO20230074 | 2023-01-26 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| WO2024158296A1 true WO2024158296A1 (en) | 2024-08-02 |
Family
ID=91970964
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/NO2024/050018 WO2024158296A1 (en) | 2023-01-26 | 2024-01-25 | A method for freeze-drying biological matter |
Country Status (2)
| Country | Link |
|---|---|
| NO (1) | NO20230074A1 (en) |
| WO (1) | WO2024158296A1 (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US2353986A (en) * | 1938-11-07 | 1944-07-18 | Sharp & Dohme Inc | Preservation |
| US20120283689A1 (en) * | 2004-10-27 | 2012-11-08 | Aseptic Technologies S.A., | Process for preparing a lyophilized material |
| US20170203871A1 (en) * | 2016-01-18 | 2017-07-20 | Vascular Solutions, Inc. | System and method for freeze-drying and packaging |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6517526B1 (en) * | 2000-12-29 | 2003-02-11 | Yehuda Tamari | Container for lyophilizing biological products |
| WO2019018195A1 (en) * | 2017-07-17 | 2019-01-24 | Baxter International Inc. | Terminal sterilization filtration closed lyophilization in a container |
| CN111295094A (en) * | 2017-10-09 | 2020-06-16 | 泰尔茂比司特生物技术有限公司 | Freeze-drying container and method for using freeze-drying container |
-
2023
- 2023-01-26 NO NO20230074A patent/NO20230074A1/en unknown
-
2024
- 2024-01-25 WO PCT/NO2024/050018 patent/WO2024158296A1/en unknown
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US2353986A (en) * | 1938-11-07 | 1944-07-18 | Sharp & Dohme Inc | Preservation |
| US20120283689A1 (en) * | 2004-10-27 | 2012-11-08 | Aseptic Technologies S.A., | Process for preparing a lyophilized material |
| US20170203871A1 (en) * | 2016-01-18 | 2017-07-20 | Vascular Solutions, Inc. | System and method for freeze-drying and packaging |
Also Published As
| Publication number | Publication date |
|---|---|
| NO20230074A1 (en) | 2024-07-29 |
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