CN110531085A - A kind of magnetic microparticle chemiluminescence detection kit and preparation method thereof measuring human nerve silk light chain protein content - Google Patents
A kind of magnetic microparticle chemiluminescence detection kit and preparation method thereof measuring human nerve silk light chain protein content Download PDFInfo
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Abstract
The invention discloses a kind of magnetic microparticle chemiluminescence kit and preparation method thereof for measuring human nerve silk light chain protein content, kit includes: neurofilament light chain protein determination R1 reagent, R2 reagent, Magneto separate reagent, calibration object liquid series and Chemoluminescent substrate.The reaction pattern for the sandwich method that the present invention uses, the principle that isolation technics combines is immunized with magnetic particle using chemiluminescence detection technology, neurofilament light chain protein content in a variety of samples such as quantitative detection human serum, blood plasma or cerebrospinal fluid, ensure the sensitivity of detection, this kit compared with conventional reagents box high sensitivity, pollution-free, high specificity, it is easy to operate and to the pre-treatment of sample require low, detectable sample type extensively, can fast high-flux detect high-volume sample, be convenient for clinical reagent application.The present invention provides a kind of more acurrate, accurate, convenient, fast and simple method for the neurofilament light chain albumen in clinical detection human serum.
Description
Technical field
Human nerve silk is detected using magnetic microparticle chemiluminescence technology and Ag-Ab combination technology the present invention relates to a kind of
Magnetic microparticle chemiluminescence detection kit of light chain protein content and preparation method thereof.The invention belongs to medical diagnosis necks
Domain.
Background technique
Neurofilament protein is the main cytoskeletal protein of neuron, respectively by neurofilament protein light chain, middle chain and heavy chain
Composition, is specifically expressed in axon and aixs cylinder.The major function of neurofilament protein is to provide structural support for aixs cylinder and regulate and control
The growth diameter of neural axon affects the rate and accuracy of neural traffic.Neurofilament protein light chain (neurofilament
Light, NFL) it is most important constituent in neurofilament protein, molecular weight 68KDa is the marker of axonal injury.Mind
Organizine light chain constitutes the skeleton of heavy chain, and heavy chain aggregation forms neural silk fiber.Due to direct wound or slowly move back
Change process, impaired nerve cell content are discharged to the compartment on periphery, it is possible to quantitative detection neuronin.It is moved back a variety of
The neurofilament of the row disease such as patient of cerebral injury, multiple sclerosis, frontotemporal dementia and other a variety of nervus retrogression diseases is light
Catenin level can be increased.
Alzheimer disease (Alzheimer's disease, AD) is a kind of Neuro-degenerative for carrying out sexual development
Disease often shows as essential characteristic with dementias such as memory disorders, personality and behavior changes in clinic, and that falls ill after 65 years old is referred to as
For Delayed onset AD.With population in the world aging, the number of AD patient is quicklyd increase, it is contemplated that the year two thousand thirty, global AD patient will
Reach 6,6,000,000 people, China will be more than 1,6,000,000 people.Compared with normal aging people, AD minimal invasive treatment's self-care ability is poor, simultaneously
It is often accompanied by mental act exception, great burden is brought to family and society, has taken over myocardial infarction, cancer, apoplexy
Summation, to society and family exert heavy pressures on.The cause of disease of the disease is unknown at present, once studies have reported that AD and cerebrospinal fluid neutralize
Neurofilament protein content in blood has certain connection, especially with light chain protein therein (neurofilamentlight,
NFL) in close relations.Therefore, research and development neurofilament light chain protein diagnostic kit has very important clinical value.
The neurofilament light chain method of protein detection being currently known is mainly enzyme-linked immunosorbent assay, fluoroimmunoassay
Method.But these methods there are still sensitivity it is low, the range of linearity is narrow, unstable result the defects of.
Therefore, it is still necessary to a kind of high sensitivity, easy to operate, high degree of automation, result stabilizations, low-cost mind at present
Organizine light chain protein detection kit and application method.
Summary of the invention
It is stable, low-cost the present invention is directed to develop a kind of high sensitivity, easy to operate, high degree of automation, result
A kind of magnetic microparticle chemiluminescence detection kit and preparation method thereof measuring human nerve silk light chain protein content.
Based on above-mentioned purpose, the present invention provides a kind of magnetic microparticle chemiluminescence for measuring human nerve silk light chain protein content
Detection kit, comprising: R1 reagent, R2 reagent, Magneto separate reagent, calibration object liquid series and Chemoluminescent substrate；Wherein, R1
Reagent is the anti-neurofilament light chain protein monoclonal antibody dilution of marked by fluorescein isothiocyanate, and R2 reagent is alkaline phosphatase
The anti-neurofilament light chain protein antibodies dilution of label, Magneto separate reagent are that anti-fluorescein isothiocynate monoclonal antibody is coated
Magnetic particle diluent, calibration object liquid series are the antigenic dilution containing various concentration neurofilament light chain albumen, chemiluminescence bottom
Thing liquid is the substrate solution of alkaline phosphatase catalytic luminescence.
Wherein, it is preferred that the R1 reagent is the different sulphur cyanogen that concentration made of being diluted as buffer is 0.3~0.9 μ g/mL
The fluorescein-labeled anti-neurofilament light chain protein monoclonal antibody dilution of acid.
Wherein, it is preferred that the pH of cushioning fluid for being used to prepare R1 reagent is 7.2~8.0, and buffer includes that concentration is 12.0
It is sodium chloride that Sodium azide that the Tris of~12.3g/L, concentration are 1.98~1.99g/L, concentration are 5.7~5.9g/L, mole dense
Spending the 0.8~1.2mL/L of magnesium chloride solution for being 1M, 0.8~1.2mL/L of liquor zinci chloridi that molar concentration is 0.1M, concentration is
The newborn bovine serum that the bovine serum albumin(BSA) and concentration that the fishskin gelatin of 5-20g/L, concentration are 2~5g/L are 10~50g/L,
Remaining is deionized water.
Wherein, it is preferred that the R2 reagent is that concentration made of being diluted as buffer is 0.5~2.0 μ g/mL alkaline phosphatase
The anti-neurofilament light chain protein antibodies dilution of enzyme label.
Wherein, it is preferred that the pH value for being used to prepare the buffer of R2 reagent is 7.2~8.0, the AP being preferably commercialized
Conjugate Stabilizer。
Wherein, it is preferred that the Magneto separate reagent is that concentration made of being diluted as buffer is that resisting for 0.5~2mg/mL is different
The coated magnetic particle solution of thiocyanic acid fluorescein monoclonal antibody.
Wherein, it is preferred that the pH value for being used to prepare the buffer of Magneto separate reagent is 7.5~9.0, and buffer composition is packet
Include the chlorine that Sodium azide, concentration that Tris, concentration that concentration is 10.5~11.3g/L are 1.91~1.95g/L are 5.5~5.7g/L
Change liquor zinci chloridi 0.8 that 0.8~1.2mL/L of magnesium chloride solution, molar concentration that sodium, molar concentration are 1M are 0.1M~
The superfine horse serum that the bovine serum albumin(BSA) and concentration that 1.2mL/L, concentration are 4.7~4.9g/L are 4.8~5.0g/L, remaining
Group is divided into deionized water.
Wherein, it is preferred that the calibration object liquid series is light containing various concentration neurofilament made of being diluted as buffer
The antigenic dilution of catenin.
Wherein, it is preferred that be used to prepare calibration object liquid series buffer include concentration be 8.5~10.2g/L Tris,
Tetracycline hydrochloride that sodium chloride that concentration is 11.8~14.5g/L, concentration are 0.008~0.028g/L, concentration is 0.008~
Casein that Sodium azide that the neomycinsulphate of 0.028g/L, concentration are 1.5~2.0g/L, concentration are 6.0~10.0g/L and
Concentration is 2.6-~3.4g/L polysorbas20, remaining group is divided into deionized water.
Wherein, it is preferred that the concentration range of the calibration object liquid series be 0~500pg/ml, pH of cushioning fluid be 7.0~
8.0。
Wherein, it is preferred that the calibration object liquid series include respectively containing concentration be 0pg/ml, 10pg/ml, 50pg/ml,
The antigenic dilution of 100pg/ml, 250pg/ml, 500pg/ml neurofilament light chain albumen.
Wherein, it is preferred that the Chemoluminescent substrate is to contain 0.2~0.4mg/mL alkali made of being diluted as buffer
The Chemoluminescent substrate of acid phosphatase catalytic luminescence substrate；It is furthermore preferred that the buffer is mole that pH value is 8~10
Concentration is the Tris-HCl buffer of 0.1-0.3M, and the alkaline phosphatase catalytic luminescence substrate is dioxane chemical combination
Object (APCL).
Wherein, it is preferred that the Chemoluminescent substrate is the Tris-HCl that the molar concentration for being 9.3 by pH value is 0.2M
The Chemoluminescent substrate of dioxane compound (APCL) containing 0.3mg/mL made of buffer dilution.
Further, the invention also provides a kind of magnetic microparticle chemiluminescences for measuring human nerve silk light chain protein content
The preparation method of detection kit, includes the following steps:
Reagent preparation R1:1) according to reagent R1 buffer composition content preparation buffer, adjust pH；2) isothiocyanic acid is prepared
Fluorescein-labeled anti-neurofilament light chain protein monoclonal antibody；3) by the anti-neurofilament light chain egg of marked by fluorescein isothiocyanate
White monoclonal antibody is diluted with R1 buffer.
Reagent preparation R2:1) according to reagent R2 buffer composition content preparation buffer, adjust pH；2) alkaline phosphatase is prepared
The anti-neurofilament light chain protein antibodies of enzyme label；3) the anti-neurofilament light chain protein antibodies of alkali phosphatase enzyme mark are buffered with R2
Liquid is diluted.
It prepares magnetic particle: 1) preparing buffer according to magnetic particle buffer composition content；2) anti-isosulfocyanic acid fluorescence is prepared
The plain coated magnetic particle of monoclonal antibody；3) magnetic particle is diluted with Tris-HCl buffer.
It prepares calibration object: 1) preparing buffer according to calibration object buffer composition content；2) neurofilament light chain albumen is resisted
Original, which is dissolved in calibration object buffer, is configured to different concentration.
It prepares Chemoluminescent substrate: 1) preparing buffer according to Chemoluminescent substrate buffer composition content；2) will
The chemiluminescent substrate of alkaline phosphatase catalytic luminescence is dissolved in buffer.
Further, the invention also provides a kind of magnetic particle chemistry hairs for measuring human nerve silk light chain protein content
Light detection kit application method, includes the following steps:
(1) by sample to be tested or 37 DEG C of calibration object, reagent R1, reagent R2 mixing incubation 15min；
(2) magnetic particle is added after above-mentioned reaction system and continues to incubate 5min in 37 DEG C；
(3) it washs, luminous substrate is added after removing unbonded antibody and impurity；
(4) luminous substrate is added, ALP catalysis substrate measures relative luminous intensity (RLU) after shining.
RLU and neurofilament light chain proteantigen concentration are proportional in a certain range, can be from standard by interpolation method
The neurofilament light chain protein content of sample to be tested is read on curve.
Methodology appraising datum when detection kit measurement human nerve silk light chain protein content above-mentioned using the present invention
It can reach following index:
Sensitivity-minimum detectable activity is 0.1pg/ml；
Neurofilament light chain albumen linear detection range, 0.1~500pg/ml；
Precision is respectively less than 8% between precision and analysis in precision-analysis, meets national requirements, illustrates examination of the present invention
Agent box has good repeatability during the experiment；
Accuracy-, which uses the serum of known neurofilament light chain protein concentration, goes hormone serum after different proportion dilutes
The rate of recovery is 95%~115%；
If the interferent concentration contained in interference-sample meet it is claimed below, on testing result without influence: bilirubin≤
400umol/L, hemoglobin≤5g/L, chyle≤0.30%, VC≤ 0.5g/L, heparin sodium≤100IU/mL.
Compared to the prior art, the beneficial effects of the present invention are:
Isolation technics is immunized using chemiluminescence detection technology and magnetic particle in the reaction pattern for the sandwich method that the present invention uses
The principle combined, the neurofilament light chain protein content in a variety of samples such as quantitative detection human serum, blood plasma or cerebrospinal fluid, it is ensured that
The sensitivity of detection, this kit compared with conventional reagents box high sensitivity (traditional ELISA kits sensitivity only~
0.1ng/ml), pollution-free, high specificity, easy to operate and require low, detectable sample type wide the pre-treatment of sample
It is general, can fast high-flux detect high-volume sample, be convenient for clinical reagent application.The present invention is the nerve in clinical detection human serum
Silk light chain protein provides a kind of more acurrate, accurate, convenient, fast and simple method.
Detailed description of the invention
Fig. 1 is concentration-luminous value curve graph of neurofilament light chain albumen in 2 kit of the embodiment of the present invention；
Specific embodiment
The invention will now be further described with reference to specific embodiments, the advantages and features of the present invention will be with description and
It is apparent.
A kind of preparation for the magnetic microparticle chemiluminescence detection kit for measuring human nerve silk light chain protein content of embodiment 1
The kit includes R1 reagent, R2 reagent, Magneto separate reagent, calibration object liquid series and chemiluminescent substrate
Liquid.
Wherein, R1 reagent includes: 1) R1 antibody: the anti-neurofilament light chain albumen list of fluorescein isothiocynate (FITC) label
Clonal antibody, concentration are 0.6 μ g/ml；2) buffer: including Tris, concentration 12.0g/L；Sodium azide, concentration 1.98g/L；
Sodium chloride, concentration 5.9g/L；Molar concentration is the magnesium chloride solution of 1M, 1.0mL/L；Molar concentration is that the zinc chloride of 0.1M is molten
Liquid, 1.0mL；Fishskin gelatin, concentration 10g/L；Bovine serum albumin(BSA), 5g/L；Newborn bovine serum, 30g/L；Remaining is deionization
Water.The pH of cushioning fluid of R1 reagent is 8.0.
Wherein, R2 reagent includes: 1) R2 antibody: the anti-neurofilament light chain protein antibodies of alkali phosphatase enzyme mark, concentration are
0.5μg/ml；2) buffer: for the AP Conjugate Stabilizer of commercialization.The pH of cushioning fluid of R2 reagent is 8.0.
Wherein, Magneto separate reagent includes: 1) magnetic particle: the coated magnetic of anti-fluorescein isothiocynate (FITC) monoclonal antibody
Particle, concentration 1mg/ml；2) buffer: including Tris, concentration 11.08g/L；Sodium azide, concentration 1.917g/L；Chlorination
Sodium, concentration 5.56g/L；Molar concentration is the magnesium chloride solution of 1M, 1.0mL/L；Molar concentration is the liquor zinci chloridi of 0.1M,
1.0mL/L；Bovine serum albumin(BSA), concentration 4.81g/L；Superfine horse serum, concentration 4.91g/L, remaining group are divided into deionization
Water.The pH of cushioning fluid of Magneto separate reagent is 8.0.
Wherein, calibration object liquid series includes: 1) neurofilament light chain proteantigen；2) buffer: including Tris, concentration is
9.1g/L；Sodium chloride, concentration 12.9g/L；Tetracycline hydrochloride, concentration 0.01g/L；Neomycinsulphate, concentration are
0.01g/L；Sodium azide, concentration 2.0g/L；Casein 10.0g/L；Polysorbas20, concentration 3.1g/L.Calibration object liquid series
PH of cushioning fluid is 7.6.Calibration object series include containing various concentration (0pg/ml, 5pg/ml, 10pg/ml, 25pg/ml,
50pg/ml, 100pg/ml) neurofilament light chain proteantigen calibration object.
Wherein, Chemoluminescent substrate is the Tris-HCl buffer dilution that the molar concentration for being 9.3 by pH value is 0.2M
Made of the dioxane compound (APCL) containing 0.3mg/mL Chemoluminescent substrate.
Embodiment 2 measures the application method of the magnetic microparticle chemiluminescence detection kit of human nerve silk light chain protein content
(1) by the calibration object of 50 μ l of embodiment 1 series (neurofilament light chain proteantigen concentration be respectively 0pg/ml,
10pg/ml, 50pg/ml, 100pg/ml, 250pg/ml, 500pg/ml) respectively with 50 μ l reagent R1, the 50 μ l reagents of embodiment 1
R2 is sequentially added in reaction tube, and mixing incubates 15min under the conditions of 37 DEG C；
(2) by mentioned reagent series respectively again in conjunction with 25 μ l Magneto separate reagents of embodiment 1 after continue to incubate in 37 DEG C
5min；
(3) 3 times are washed to remove unbonded antibody and impurity with cleaning solution；
(4) 150 μ l, the ALP catalysis substrate of luminous substrate liquid of addition embodiment 1 is surveyed after shining using chemiluminescence detector
Determine relative luminous intensity (RLU), as a result as shown in table 1 below:
Table 1
| Calibration object (pg/ml) | 0 | 10 | 50 | 100 | 250 | 500 |
| RLU(100000) | 0.013 | 1.419 | 6.178 | 12.238 | 28.562 | 54.967 |
(5) it is fitted according to the numerical value of table 1, it is bent to obtain neurofilament light chain protein concentration shown in FIG. 1-luminous value standard
Line.
(6) 50 μ l reagent R1, the 50 μ l reagent R2 of the test serum sample of 50 μ l and embodiment 1 are sequentially added into reaction tube
In, mixing incubates 15min under the conditions of 37 DEG C；
(7) by mentioned reagent again in conjunction with 25 μ l Magneto separate reagents of embodiment 1 after continue in 37 DEG C incubate 5min；
(8) (contain 0.02% polysorbas20 and the 0.1M of sodium chloride of 15w/w%, the Tris- that pH value is 8 with cleaning solution
HCl buffer.) washing 3 times antibody and impurity unbonded with removal；
(9) 150 μ l, the ALP catalysis substrate of luminous substrate liquid of addition embodiment 1 is surveyed after shining using chemiluminescence detector
Determining relative luminous intensity RLU is 142000；
(10) according to the neurofilament light chain protein concentration of step (5)-luminous value standard curve calculate RLU be 142000 to
Surveying the corresponding neurofilament light chain protein concentration values of blood serum sample is 10pg/ml.
The magnetic microparticle chemiluminescence detection kit of human nerve silk light chain protein content provided by the invention can with it is complete
The combination of robotics luminescence analyzer, operating procedure greatly simplify, and increase detection speed and detection flux, improve detection effect
Rate, while avoiding error caused by manual operation.
The experiment of the Sensitivity comparison of comparative example kit of the invention and traditional ELISA kits
Both kit and traditional ELISA kits of the invention are used respectively, and same sample is detected, determine
Sensitivity, as a result as shown in table 2 and table 3:
The kit sensitivity of the present invention of table 2
3 traditional ELISA kits sensitivity of table
Kit and traditional ELISA kits of the invention are used respectively, to CSF sample, serum sample and blood plasma
Sample is detected, and the results are shown in Table 4:
The kit of the present invention of table 4 and traditional ELISA kits pattern detection comparison data
The above result shows that this kit high sensitivity, detectable sample type compared with traditional ELISA kits is wide
It is general, can fast high-flux detect high-volume sample, be convenient for clinical reagent application.
It should be understood by those ordinary skilled in the art that: the discussion of any of the above embodiment is exemplary only, not
It is intended to imply that the scope of the present invention (including claim) is limited to these examples；Under thinking of the invention, above embodiments
Or it can also be combined between the technical characteristic in different embodiments, and there are different aspects present invention as described above
Many other variations, in order to it is concise they do not provided in details.Therefore, all within the spirits and principles of the present invention,
Any omission, modification, equivalent replacement, improvement for being made etc., should all be included in the protection scope of the present invention.
Claims (10)
1. a kind of magnetic microparticle chemiluminescence detection kit for measuring human nerve silk light chain protein content, it is characterised in that: packet
It includes: R1 reagent, R2 reagent, Magneto separate reagent, calibration object liquid series and Chemoluminescent substrate；
Wherein, R1 reagent is the anti-neurofilament light chain protein monoclonal antibody dilution of marked by fluorescein isothiocyanate, R2 reagent
For the anti-neurofilament light chain protein antibodies dilution of alkali phosphatase enzyme mark, Magneto separate reagent is anti-fluorescein isothiocynate Dan Ke
The grand coated magnetic particle diluent of antibody, calibration object liquid series are the antigen diluent containing various concentration neurofilament light chain albumen
Liquid, Chemoluminescent substrate are the substrate solution of alkaline phosphatase catalytic luminescence.
2. kit as described in claim 1, it is characterised in that: the R1 reagent is that concentration made of being diluted as buffer is
The anti-neurofilament light chain protein monoclonal antibody dilution of the marked by fluorescein isothiocyanate of 0.3~0.9 μ g/mL；
Wherein, it is preferred that be used to prepare R1 reagent pH of cushioning fluid be 7.2~8.0, buffer include concentration be 12.0~
Sodium chloride that Sodium azide that the Tris of 12.3g/L, concentration are 1.98~1.99g/L, concentration are 5.7~5.9g/L, molar concentration
0.8~1.2mL/L of liquor zinci chloridi, the concentration 5- for being 0.1M for 0.8~1.2mL/L of magnesium chloride solution of 1M, molar concentration
The newborn bovine serum that the bovine serum albumin(BSA) and concentration that the fishskin gelatin of 20g/L, concentration are 2~5g/L are 10~50g/L,
Remaining is deionized water.
3. kit as described in claim 1, it is characterised in that: the R2 reagent is that concentration made of being diluted as buffer is
The anti-neurofilament light chain protein antibodies dilution of 0.5~2.0 μ g/mL alkali phosphatase enzyme mark；
Wherein, it is preferred that the pH value for being used to prepare the buffer of R2 reagent is 7.2~8.0, the AP being more preferably commercialized
Conjugate Stabilizer。
4. kit as described in claim 1, it is characterised in that: the Magneto separate reagent is dense made of being diluted as buffer
Degree is the coated magnetic particle solution of anti-fluorescein isothiocynate monoclonal antibody of 0.5~2mg/mL；
Wherein, it is preferred that be used to prepare the buffer of Magneto separate reagent pH value be 7.5~9.0, buffer composition be include dense
Spend the chlorination that the Tris for being 10.5~11.3g/L, the Sodium azide that concentration is 1.91~1.95g/L, concentration are 5.5~5.7g/L
0.8~1.2mL/ of liquor zinci chloridi that 0.8~1.2mL/L of magnesium chloride solution that sodium, molar concentration are 1M, molar concentration are 0.1M
L, the superfine horse serum that the bovine serum albumin(BSA) and concentration that concentration is 4.7~4.9g/L are 4.8~5.0g/L, remaining group are divided into
Deionized water.
5. kit as described in claim 1, it is characterised in that: the calibration object liquid series is made of being diluted as buffer
Antigenic dilution containing various concentration neurofilament light chain albumen；
Wherein, it is preferred that it is the Tris of 8.5~10.2g/L, concentration that the buffer for being used to prepare calibration object liquid series, which includes concentration,
For the sodium chloride of 11.8~14.5g/L, concentration be 0.008~0.028g/L tetracycline hydrochloride, concentration be 0.008~
Casein that Sodium azide that the neomycinsulphate of 0.028g/L, concentration are 1.5~2.0g/L, concentration are 6.0~10.0g/L and
Concentration is the polysorbas20 of 2.6-~3.4g/L, remaining group is divided into deionized water.
6. kit as claimed in claim 5, it is characterised in that: the concentration range of the calibration object liquid series be 0~
500pg/ml, pH of cushioning fluid are 7.0~8.0；
Wherein, it is preferred that the calibration object liquid series include respectively containing concentration be 0pg/ml, 10pg/ml, 50pg/ml,
The antigenic dilution of 100pg/ml, 250pg/ml, 500pg/ml neurofilament light chain albumen.
7. kit as described in claim 1, it is characterised in that: the Chemoluminescent substrate is diluted by buffer
The Chemoluminescent substrate containing 0.2~0.4mg/mL alkaline phosphatase catalytic luminescence substrate；
Wherein, it is preferred that the buffer is the Tris-HCl buffering that the molar concentration that pH value is 8~10 is 0.1-0.3M
Liquid, the alkaline phosphatase catalytic luminescence substrate are dioxane compound (APCL).
8. kit as claimed in claim 7, it is characterised in that: it is 9.3 to rub that the Chemoluminescent substrate, which is by pH value,
Dioxane compound (APCL) containing 0.3mg/mL made of the Tris-HCl buffer dilution that your concentration is 0.2M
Chemoluminescent substrate.
9. the described in any item kits of claim 1-8 neurofilament light chain protein content reagent in preparation measurement human sample
In purposes.
10. purposes as claimed in claim 9, it is characterised in that: the sample is human serum, blood plasma or cerebrospinal fluid.
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| CN116675768A (en) * | 2023-05-10 | 2023-09-01 | 武汉爱博泰克生物科技有限公司 | Rabbit monoclonal antibody aiming at human NEFL protein, and preparation method and application thereof |
| CN117233404A (en) * | 2023-11-13 | 2023-12-15 | 北京豪迈生物工程股份有限公司 | Kit for determining protein 2 content in field of human coiled coil structure |
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