CN109988221A - A kind of preparation method of glycoprotein quality-control product - Google Patents
A kind of preparation method of glycoprotein quality-control product Download PDFInfo
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- CN109988221A CN109988221A CN201910271440.3A CN201910271440A CN109988221A CN 109988221 A CN109988221 A CN 109988221A CN 201910271440 A CN201910271440 A CN 201910271440A CN 109988221 A CN109988221 A CN 109988221A
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Abstract
The present invention relates to quality-control product preparation fields, in particular to a kind of preparation method of glycoprotein quality-control product, comprising the following steps: glycoprotein solution to be separated is reacted with perchloric acid, obtains reaction solution；The reaction solution carries out break process, is then isolated to the solution containing glycoprotein；Solution containing the glycoprotein is dialysed to obtain the final product.The preparation method of glycoprotein quality-control product provided by the invention can effectively promote the yield of destination protein, and repeatability and stability are good.
Description
Technical field
The present invention relates to quality-control product preparation fields, in particular to a kind of preparation method of glycoprotein quality-control product.
Background technique
External diagnosis reagent is to carry out vitro detection simultaneously by the sample (tissue, body fluid, blood, metabolin etc.) to human body
Obtain the product of clinical assistant diagnosis information.Currently, China's external diagnosis reagent market scale is about 5,000,000,000 or so, Nian Zeng every year
Long rate is 20%~30%, and significantly larger than the growth rate of the developed countries such as America and Europe 12%, domestic external diagnosis reagent are one
A huge market.Clinically there are many demand for tumour diagnostic reagent, are the staple products of many diagnostic reagent manufacturers,
So markers in diagnosis raw material market is wide, trans-corporation occupies most of share in the market.
Tumor markers (Tumor Marker) are chemicals existing for reflection tumour, they or be not present in normal
Adult tissue and be detected in embryonic tissue, or content in tumor tissues substantially exceeds the content in normal tissue, they
Presence or quantitative change can prompt the property of tumour, so as to understanding tissue generations, the cell differentiation, cell function of tumour, to help
Help diagnosis, classification, Index for diagnosis and the treatment guidance of tumour.Sugar antigen (carbohydrate antigen, CA) is swollen
A major class in tumor markers, generally includes CA50, CA125, CA724, CA199, CA153 etc..
Quality-control product is used exclusively for the sample or solution of quality control purpose, cannot act as calibrating.It is to make user
The use situation of detection system can be understood in time and additionally configured.Basic material used in quality-control product is prepared generally to come from
The serum of human or animal also uses other body fluid sometimes.Consider that quality-control product is preferably from human body from matrix otherness.
For general quality-control product, stability is its most important index.Good quality-control product can save 1 under defined preservation condition
By 2 years.In addition, the specificity of quality-control product is very strong, it is often only provided with the definite value of company.Therefore, for routine quality control
Quality-control product does not all need the trackability for yet having definite value.The famous Bio-rad company of production quality-control product declares its public affairs
It takes charge of all quality-control products and does not have the trackability on measuring, and this also complies with the requirement of ISO 17511.
Summary of the invention
The purpose of the present invention is to provide a kind of preparation methods of glycoprotein quality-control product.
In order to realize above-mentioned purpose of the invention, the following technical scheme is adopted:
In a first aspect, a kind of preparation method of glycoprotein quality-control product provided by the invention, comprising the following steps:
Glycoprotein solution to be separated is reacted with perchloric acid, obtains reaction solution；
The reaction solution carries out break process, is then isolated to the solution containing glycoprotein；
Solution containing the glycoprotein is dialysed to obtain the final product.
The preparation method of glycoprotein quality-control product provided by the invention, compared to the preparation of existing non-quality-control product glycoprotein
Method, the step of increasing break process, this is because the present inventor has found in actual operation, glycoprotein solution to be separated with
In perchloric acid reaction process, spawn is generated, and after break process, the ingredient of reaction solution can be made to disperse, be convenient for
The separation of destination protein effectively improves the yield of destination protein.
Second aspect, the present invention also increase following steps before perchloric acid processing:
To the glycoprotein solution to be separated first in -20 DEG C of ± 5 DEG C of pre-cooling 10-25min, then reacted with perchloric acid.It should
Step is further obviously improved the yield of destination protein.
The third aspect, it is a discovery of the invention that the glycoprotein solution obtained after perchloric acid purifies, without adding alkali neutralization, yield is anti-
And it improves.
The optional sugar antigen of glycoprotein in the present invention, appointing preferably in CA50, CA125, CA724, CA199 and CA153
It is a kind of.
Compared with prior art, the invention has the benefit that
(1) present invention increases after glycoprotein solution to be separated is reacted with perchloric acid carries out broken step to reaction solution,
Effectively improve the yield of glycoprotein.
(2) present invention increases before glycoprotein solution to be separated is reacted with perchloric acid to -20 DEG C of glycoprotein solution to be separated
± 5 DEG C of pretreatments, effectively improve the yield of glycoprotein.
(3) it has also been found that, without adding alkali neutralization, yield mentions the glycoprotein solution obtained after perchloric acid purifies instead
It is high.
Specific embodiment
Perchloric acid is the oxyacid of chlorine, is acid strongest in inorganic oxacid.It can cause albuminous degeneration to solidify, and
And for many albumen, this solidification is irreversible, and glycoprotein has acid resistance, therefore, it is possible to use perchloric acid
It is denaturalized other albumen therein and achievees the purpose that purified glycoproteins.
In existing method, has the relevant preparation method of sugar antigen, such as ovarian cancer resistance related antigen monoclonal antibody
Preparation and immunoassay method foundation, CA125 related antigen is purified in seroperitoneum, referring to the method for Davis etc., through height
Chloric acid separation, gel chromatography and Affi-Gel indigo plant column chromatographic purifying obtain carrying the Ovarian Cancer Associated Antigen of CA125 antigenic determinant.
A kind of for another example foundation of the tube-type chemical luminous detection method for oophoroma related neoplasms marker CA125,
The extraction of CA125 native antigen and purification process, comprising: perchloric acid is added under condition of ice bath in ascites centrifugal filtration, precipitates
30min, subsequent 12000rpm centrifugation 15min collect supernatant, are added after NaOH is neutralized and supernatant is collected by centrifugation, carry out to supernatant saturating
Analysis.
But it finding in practical applications, existing method can not adapt to the preparation demand of glycoprotein quality-control product in the present invention,
It is mainly shown as that yield is low, stability is bad.
In view of the above problems, the present invention provides a kind of preparation method of glycoprotein quality-control product, comprising the following steps:
Glycoprotein solution to be separated is reacted with perchloric acid, obtains reaction solution；
The reaction solution carries out break process, is then isolated to the solution containing glycoprotein；
Solution containing the glycoprotein is dialysed to obtain the final product.
The preparation method of glycoprotein quality-control product provided by the invention, compared to the preparation side of existing non-quality-control product glycoprotein
Method, the step of increasing break process, this is because the present inventor has found in actual operation, glycoprotein solution to be separated and high
In chloric acid reaction process, spawn is generated, and after break process, the ingredient of reaction solution can be made to disperse, be convenient for mesh
Albumen separation, effectively improve the yield of destination protein.
In the present invention, broken means can be to be a variety of, as ultrasonication, attrition crushing, shearing-crushing, stirring are broken.
Preferably, described broken using ultrasonication.By ultrasonication, the substance in gelatinous reaction solution obtains one
Determine the dispersion of degree, so that the glycoprotein wherein embedded exposes, is convenient for subsequent separation.
Preferably, the power of the ultrasonication is 13 ± 3W, the time of the ultrasonication is 1-5min, preferably
2-3min, by the program treated reaction solution after subsequent processing, the yield of glycoprotein is substantially improved.Wherein, ultrasonic
Mode can be continous way, be also possible to it is intermittent, it is intermittent to take lasting 2-4s, rest 2-4s
Further, the mass fraction of the perchloric acid solution is 60%-72%.Such as in various embodiments, high chlorine
The mass concentration of acid solution can be 60%, 62%, 63%, 65%, 68%, 70%, 72% etc..
In the present invention, in principle, the concentration of the glycoprotein to be separated in glycoprotein solution to be separated is not limited, generally,
The concentration of glycoprotein to be separated used is 1000IU/mL or more.It such as can be 1000IU/mL, 1100IU/mL, 1200IU/
ML, 1300IU/mL, 1400IU/mL, 1500IU/mL, 1600IU/mL, 2000IU/mL, 2500IU/mL, 3000IU/mL etc..
Further, the volume ratio of the glycoprotein solution to be separated and the perchloric acid solution is 3-5mL:10-100 μ
L。
Preferably, the volume ratio of the glycoprotein solution to be separated and the perchloric acid solution is 4mL:25-75 μ L；Though
So the volume of the perchloric acid solution of different glycoprotein addition is slightly different, but perchloric acid solution additive amount energy in 25-75 μ L
Obtain the effect that albumen yield is higher than 45%.
Preferably, the glycoprotein solution to be separated is first in -20 DEG C of ± 5 DEG C of pre-cooling 10-25min, then with perchloric acid
Reaction.
It is a discovery of the invention that glycoprotein solution to be separated increases the step of -20 DEG C of ± 5 DEG C of pre-cooling 10-25min, then use
Perchloric acid extracts glycoprotein, hence it is evident that increases the yield of glycoprotein.
In the present invention, glycoprotein solution to be separated is during pre-cooling, the state being kept stirring, and solution holding is not tied
Ice is in a liquid state.
Further, perchloric acid solution is added dropwise at 0 ± 2 DEG C in the sample after pre-cooling while stirring.
Glycoprotein solution to be separated involved in the present invention is not limited to cell fermentation liquid, is also possible to glycoprotein containing purpose
Tissue fluid.
Further, the glycoprotein solution to be separated be cell fermentation liquid when, can it is concentrated, separate, be obtained by filtration；
The filtering can be carried out successively using qualitative filter paper and 0.22 μm of filter membrane, to remove impurity.
Isolated purpose is by the impurity removal in reaction solution, therefore, selection mode or parameter according to actual needs.
Further, the separation is carried out by the way of centrifugation；
Further, the revolution of the centrifugation is 10000-12000rpm, and the temperature of centrifugation is 0-15 DEG C, the time of centrifugation
For 3-20min.Such as, the temperature of centrifugation can be 0 DEG C, 4 DEG C, 8 DEG C, 10 DEG C, 12 DEG C, 15 DEG C etc.；The time of centrifugation can be
3min, 5min, 8min, 10min, 15min, 20min etc..
Further, the dialysis is carried out using the buffer of 80-100 times of volume, can such as use chlorine containing 140-150mM
The PB buffer for changing sodium carries out, and the pH of the PB buffer is 7.0-7.5.
Further, the dialysis carries out under 4 ± 2 DEG C of environment.
Further, the time of the dialysis is 8-12h.
Further, the optional sugar antigen of the glycoprotein, preferably in CA50, CA125, CA724, CA199 and CA153
It is any.
Embodiment of the present invention is described in detail below in conjunction with embodiment, but those skilled in the art will
Understand, the following example is merely to illustrate the present invention, and is not construed as limiting the scope of the invention.It is not specified in embodiment specific
Condition person carries out according to conventional conditions or manufacturer's recommended conditions.Reagents or instruments used without specified manufacturer is
The conventional products that can be obtained by commercially available purchase.
Embodiment 1
By taking the cell fermentation liquid containing CA50 as an example, perform the following operation:
1, perchloric acid purifies
1.1 take sample to be pre-chilled 20 minutes at -20 DEG C；
1.2 in ice-water bath, and it is 70% that 50 μ L mass concentrations are slowly added dropwise into the 4mL concentrated broth being stirred continuously
Perchloric acid solution.
After 1.3 completion of dropwise addition, ultrasonication 2min (power 13W, 2.5s/2.5s).
1.4 centrifugations: 12000rpm, 4 DEG C, 5min.
1.5 take supernatant, are dialysed under 4 DEG C of environment with the PB buffer (sodium chloride containing 150mM, pH7.4) of 100 times of volumes
Night.
2, the concentration of purification of samples is measured
Purification of samples is diluted 10 times, with sugar antigen CA50 detection kit (Zhengzhou Antu biology) detection purification of samples
Concentration, purification of samples concentration are 763IU/mL.
3, the preparation of quality-control product
Purification of samples is diluted to 10IU/mL and 100IU/mL with the PB buffer of 20% people's feminine gender blood (seracon).
4, quality-control product repeated experiment
It detects two quality-control product samples each 10 times, calculates the coefficient of variation (CV).The coefficient of variation < 5%.
The repeated experiment effect of 1 quality-control product of table
5, quality-control product stability experiment
As it can be seen from table 1 after two quality-control products are placed in 37 DEG C of incubations 7 days, year-on-year 4 DEG C of storing samples signal values, letter
Number different < 15% of value difference.As it can be seen that the stability of quality-control product provided by the invention meets the requirement of quality-control product.
2 stability test result of table
Embodiment 2
By taking the cell fermentation liquid containing CA125 as an example, perform the following operation:
1.1 samples are pre-chilled 15 minutes at -20 DEG C；
1.2 in ice-water bath, and it is 70% that 50 μ L mass concentrations are slowly added dropwise into the 4mL concentrated broth being stirred continuously
Perchloric acid solution.
After 1.3 completion of dropwise addition, ultrasonication 3min (power 13W).
1.4 centrifugations: 12000rpm, 4 DEG C, 5min.
1.5 take supernatant, are dialysed under 4 DEG C of environment with the PB buffer (sodium chloride containing 150mM, pH7.4) of 100 times of volumes
Night.
Carbohydrate Antigen CA125 detection kit is used to measure above-mentioned gained sample concentration as 822IU/mL, calculating yield is
45%.Quality-control product repeated experiment measures, the coefficient of variation (CV) < 5%, and after quality-control product is placed in 37 DEG C of incubations 7 days, year-on-year 4 DEG C are deposited
Setting-out product signal value, signal value difference < 15%, meets the requirement of quality-control product.
Embodiment 3
By taking the cell fermentation liquid containing CA125 as an example, perform the following operation:
The 1.1 cell fermentation liquid containing CA125 are pre-chilled 25 minutes at -20 DEG C；
1.2 in ice-water bath, and it is 70% that 25 μ L mass concentrations are slowly added dropwise into the 4mL concentrated broth being stirred continuously
Perchloric acid solution.
After 1.3 completion of dropwise addition, ultrasonication 4min (power 10W).
1.4 centrifugations: 12000rpm, 4 DEG C, 5min.
1.5 take supernatant, are dialysed under 4 DEG C of environment with the PB buffer (sodium chloride containing 150mM, pH7.4) of 100 times of volumes
Night.
Carbohydrate Antigen CA125 detection kit is used to measure above-mentioned gained sample concentration as 949IU/mL, calculating yield is
50%.Quality-control product repeated experiment measures, the coefficient of variation (CV) < 5%, and after quality-control product is placed in 37 DEG C of incubations 7 days, year-on-year 4 DEG C are deposited
Setting-out product signal value, signal value difference < 15%, meets the requirement of quality-control product.
Embodiment 4
On the basis of embodiment 1, specific as follows using different processing modes:
1) unlike the first embodiment, by 1.1 the step of, is changed to 2-8 DEG C and handles 20 minutes, obtains sample A；
2) precooling step for unlike the first embodiment, removing 1.1 obtains sample B；
3) sample C is obtained by 1 method of embodiment；
Sample is purified according still further to the method for embodiment 1, respectively obtains sample A, B and C.
It is as shown in table 3 to measure sample concentration calculating yield.
Influence of 3 Different treatments of table to purification yield
| Sample | Concentration (IU/mL) | Volume (mL) | Yield |
| A | 277 | 182 | 21% |
| B | 321 | 188 | 25% |
| C | 865 | 189 | 68% |
Embodiment 5
On the basis of embodiment 1, specific as follows using different processing modes:
1) sample D is obtained by 1 method of embodiment；
2) unlike the first embodiment, the broken step of removal 1.3, obtains sample E:
3) unlike the first embodiment, after step 1.4 centrifugation, after taking supernatant, 10M KOH is added dropwise into supernatant, until pH
Balance is finally dialysed under 4 DEG C of environment with the PB buffer (sodium chloride containing 150mM, pH7.4) of 100 times of volumes again to 7.0-8.0
Overnight.Obtain sample F.
Concentration is measured to sample D, E and F respectively and calculates yield, the results are shown in Table 4.
The yield of the different samples of table 4
| Sample | Concentration (IU/mL) | Volume (mL) | Yield |
| D | 768 | 3.2 | 68% |
| E | 441 | 3.4 | 35% |
| F | 682 | 3.8 | 57% |
Inventors have found that the concentrate of fermentation liquid is added in perchloric acid reaction process, colloid substance will form, due to reaction
Protein cohesion is more serious in the process, will cause the embedding of target protein, using the method dispersion sol liquid of ultrasound, can drop
Low target albumen is embedded, to improve yield.
In addition, the glycoprotein solution that result above it is known that using method of the invention, obtains after perchloric acid purifies
Without adding alkali neutralization, yield improves instead.
Embodiment 6
By taking CA50, CA724, CA199 and CA153 as an example.
Sample is pre-chilled 20 minutes at -20 DEG C ± 5；In ice-water bath, slowly into the 4mL concentrated broth being stirred continuously
The perchloric acid of 25 μ L (sample 1#), 50 μ L (2#) and 75 μ L (3#) concentration 70% is added dropwise；After completion of dropwise addition, 5 are stirred in ice-water bath
Minute；Ultrasonication 2min (15W)；Centrifugation: 12000rpm, 4 DEG C, 5min；Supernatant is taken, (is contained with the PB buffer of 100 times of volumes
150mM sodium chloride, pH7.4) dialysed overnight under 4 DEG C of environment.
To different purification of samples measurement concentration and calculate yield.
1, three purification of samples of CA724 measure concentration and calculate yield, and the results are shown in Table 5.
The yield situation of three purification of samples of 5 CA724 of table
| Sample | Concentration (IU/mL) | Volume (mL) | Yield |
| CA724-3# | 677 | 3.8 | 47% |
| CA724-2# | 884 | 3.6 | 58% |
| CA724-1# | 1022 | 3.5 | 65% |
2, three purification of samples of CA50 measure concentration and calculate yield, and the results are shown in Table 6.
The yield situation of three purification of samples of 6 CA50 of table
| Sample | Concentration (IU/mL) | Volume (mL) | Yield |
| CA50-3# | 665 | 3.6 | 53% |
| CA50-2# | 843 | 3.8 | 71% |
| CA50-1# | 903 | 3.8 | 76% |
3, three purification of samples of CA199 measure concentration and calculate yield, and the results are shown in Table 7.
The yield situation of three purification of samples of 7 CA199 of table
| Sample | Concentration (IU/mL) | Volume (mL) | Yield |
| CA199-3# | 1954 | 3.6 | 74% |
| CA199-2# | 1833 | 3.5 | 67% |
| CA199-1# | 1921 | 3.8 | 77% |
4, three purification of samples of CA153 measure concentration and calculate yield, and the results are shown in Table 8.
The yield situation of three purification of samples of 8 CA153 of table
Although illustrate and describing the present invention with specific embodiment, it will be appreciated that without departing substantially from of the invention
Many other change and modification can be made in the case where spirit and scope.It is, therefore, intended that in the following claims
Including belonging to all such changes and modifications in the scope of the invention.
Claims (10)
1. a kind of preparation method of glycoprotein quality-control product, which comprises the following steps:
Glycoprotein solution to be separated is reacted with perchloric acid, obtains reaction solution；
The reaction solution carries out break process, is then isolated to the solution containing glycoprotein；
Solution containing glycoprotein is dialysed to obtain the final product.
2. preparation method according to claim 1, which is characterized in that the mass fraction of the perchloric acid solution is 60%-
72%.
3. preparation method according to claim 2, which is characterized in that the glycoprotein solution to be separated and the perchloric acid
The volume ratio of solution is 3-5mL:10-100 μ L.
4. preparation method according to claim 3, which is characterized in that the glycoprotein solution to be separated and the perchloric acid
The volume ratio of solution is 4mL:25-75 μ L.
5. preparation method according to claim 1, which is characterized in that described to be crushed including ultrasonication, attrition crushing, cut
Cut through it is broken, stirring be crushed it is any one or more of.
6. preparation method according to claim 1, which is characterized in that described to be broken for ultrasonication.
7. preparation method according to claim 1, which is characterized in that the glycoprotein solution to be separated is first at -20 DEG C ± 5
DEG C pre-cooling 10-25min, then reacted with perchloric acid.
8. preparation method according to claim 1-7, which is characterized in that the glycoprotein is sugar antigen.
9. preparation method according to claim 1-7, which is characterized in that the glycoprotein be selected from CA50,
CA125, CA724, CA199 and CA153.
10. the glycoprotein quality-control product obtained using any one of the claim 1-9 preparation method is preparing associated glycoprotein inspection
Purposes in test agent box.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106153607A (en) * | 2016-08-23 | 2016-11-23 | 江苏出入境检验检疫局动植物与食品检测中心 | A kind of clenobuterol hydrochloride chemoluminescence method quantitative determination reagent kit |
| WO2018132022A1 (en) * | 2017-01-14 | 2018-07-19 | Sds Optic Sp. Z O.O. | Device for detecting and/or determining the concentration of an analyte present in a tissue and a method and use of this device |
| CN109030638A (en) * | 2017-06-08 | 2018-12-18 | 安琪酵母股份有限公司 | A method of for measuring selenomethionine in bio-matrix |
-
2019
- 2019-04-04 CN CN201910271440.3A patent/CN109988221A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106153607A (en) * | 2016-08-23 | 2016-11-23 | 江苏出入境检验检疫局动植物与食品检测中心 | A kind of clenobuterol hydrochloride chemoluminescence method quantitative determination reagent kit |
| WO2018132022A1 (en) * | 2017-01-14 | 2018-07-19 | Sds Optic Sp. Z O.O. | Device for detecting and/or determining the concentration of an analyte present in a tissue and a method and use of this device |
| CN109030638A (en) * | 2017-06-08 | 2018-12-18 | 安琪酵母股份有限公司 | A method of for measuring selenomethionine in bio-matrix |
Non-Patent Citations (1)
| Title |
|---|
| 王旭亮: "一种针对卵巢癌相关肿瘤标志物CA125的管式化学发光检测方法的建立", 《万方中国学位论文全文数据库》 * |
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