CN111378032A - Preparation method of egg yolk antibody for clinical diagnostic reagent - Google Patents
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- 210000002969 egg yolk Anatomy 0.000 title claims abstract description 94
- 235000013345 egg yolk Nutrition 0.000 title claims abstract description 48
- 102000002322 Egg Proteins Human genes 0.000 title claims abstract description 46
- 108010000912 Egg Proteins Proteins 0.000 title claims abstract description 46
- 239000003153 chemical reaction reagent Substances 0.000 title claims abstract description 34
- 238000002360 preparation method Methods 0.000 title claims abstract description 8
- 238000000034 method Methods 0.000 claims abstract description 31
- 239000007788 liquid Substances 0.000 claims abstract description 30
- 238000001914 filtration Methods 0.000 claims abstract description 16
- 238000000108 ultra-filtration Methods 0.000 claims abstract description 13
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 11
- 238000004108 freeze drying Methods 0.000 claims abstract description 9
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims abstract description 5
- 229910021485 fumed silica Inorganic materials 0.000 claims abstract description 5
- 238000000746 purification Methods 0.000 claims abstract description 5
- 238000003759 clinical diagnosis Methods 0.000 claims abstract description 3
- 239000002671 adjuvant Substances 0.000 claims description 27
- 241000287828 Gallus gallus Species 0.000 claims description 16
- 238000001514 detection method Methods 0.000 claims description 12
- 239000006228 supernatant Substances 0.000 claims description 12
- 239000000427 antigen Substances 0.000 claims description 11
- 102000036639 antigens Human genes 0.000 claims description 11
- 108091007433 antigens Proteins 0.000 claims description 11
- 239000000243 solution Substances 0.000 claims description 10
- 235000013601 eggs Nutrition 0.000 claims description 9
- 230000036039 immunity Effects 0.000 claims description 9
- 239000008213 purified water Substances 0.000 claims description 9
- 238000005728 strengthening Methods 0.000 claims description 9
- 238000010790 dilution Methods 0.000 claims description 8
- 239000012895 dilution Substances 0.000 claims description 8
- 238000005238 degreasing Methods 0.000 claims description 7
- 239000012528 membrane Substances 0.000 claims description 7
- 238000002156 mixing Methods 0.000 claims description 7
- 210000003205 muscle Anatomy 0.000 claims description 7
- 229920001817 Agar Polymers 0.000 claims description 5
- 239000008272 agar Substances 0.000 claims description 5
- 210000000481 breast Anatomy 0.000 claims description 5
- 238000005352 clarification Methods 0.000 claims description 5
- 238000005516 engineering process Methods 0.000 claims description 5
- 230000000951 immunodiffusion Effects 0.000 claims description 5
- 239000000463 material Substances 0.000 claims description 5
- 230000020477 pH reduction Effects 0.000 claims description 5
- 239000000843 powder Substances 0.000 claims description 5
- 238000009777 vacuum freeze-drying Methods 0.000 claims description 5
- 238000007865 diluting Methods 0.000 claims description 4
- 150000002632 lipids Chemical class 0.000 claims description 4
- 239000011148 porous material Substances 0.000 claims description 4
- 238000007710 freezing Methods 0.000 claims description 3
- 230000008014 freezing Effects 0.000 claims description 3
- 230000003053 immunization Effects 0.000 claims description 3
- 238000003756 stirring Methods 0.000 claims description 3
- 102000004127 Cytokines Human genes 0.000 claims description 2
- 108090000695 Cytokines Proteins 0.000 claims description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 2
- 238000001816 cooling Methods 0.000 claims description 2
- 239000000839 emulsion Substances 0.000 claims description 2
- 239000000706 filtrate Substances 0.000 claims description 2
- 230000002163 immunogen Effects 0.000 claims description 2
- 230000000813 microbial effect Effects 0.000 claims description 2
- 102000039446 nucleic acids Human genes 0.000 claims description 2
- 108020004707 nucleic acids Proteins 0.000 claims description 2
- 150000007523 nucleic acids Chemical class 0.000 claims description 2
- 239000002245 particle Substances 0.000 claims description 2
- 241000124008 Mammalia Species 0.000 abstract description 7
- 230000008901 benefit Effects 0.000 abstract description 5
- 239000002994 raw material Substances 0.000 abstract description 5
- 230000035945 sensitivity Effects 0.000 abstract description 3
- 230000007613 environmental effect Effects 0.000 abstract 1
- 235000013330 chicken meat Nutrition 0.000 description 12
- 239000000047 product Substances 0.000 description 8
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 4
- 241001494479 Pecora Species 0.000 description 4
- 210000002966 serum Anatomy 0.000 description 4
- 241000282414 Homo sapiens Species 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 239000002504 physiological saline solution Substances 0.000 description 3
- 230000001954 sterilising effect Effects 0.000 description 3
- 238000004659 sterilization and disinfection Methods 0.000 description 3
- 108010074051 C-Reactive Protein Proteins 0.000 description 2
- 102100032752 C-reactive protein Human genes 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 230000037452 priming Effects 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 238000003908 quality control method Methods 0.000 description 2
- 241000283707 Capra Species 0.000 description 1
- 102000004856 Lectins Human genes 0.000 description 1
- 108090001090 Lectins Proteins 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 238000010241 blood sampling Methods 0.000 description 1
- 230000004154 complement system Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 239000002523 lectin Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 238000007817 turbidimetric assay Methods 0.000 description 1
- 238000004879 turbidimetry Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/02—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies from eggs
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/10—Immunoglobulins specific features characterized by their source of isolation or production
- C07K2317/11—Immunoglobulins specific features characterized by their source of isolation or production isolated from eggs
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Immunology (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention discloses a preparation method of a yolk antibody for a clinical diagnostic reagent, belonging to the field of biological antibody preparations. The invention collects qualified egg yolk, dilutes and acidifies the egg yolk, then thoroughly degreases the acidified egg yolk liquid by adding fumed silica, and further prepares the egg yolk antibody for the freeze-dried clinical diagnostic reagent by deep filtration, ultrafiltration purification and concentration and freeze drying processes. The freeze-dried yolk antibody produced by the method is an excellent substitute raw material of a mammal antibody for clinical diagnosis, and has the characteristics of easy water solubility, high sensitivity, convenient and safe use. The method has the advantages of simple process, environmental friendliness, easiness in operation, high product safety and the like.
Description
Technical Field
The invention relates to the field of biological antibody preparations, in particular to a preparation method of a yolk antibody for a clinical diagnostic reagent.
Background
The yolk antibody IgY is an immune active substance extracted from yolk, and after a chicken is immunized by a specific antigen, a specific yolk antibody is accumulated in the yolk at high concentration, extracted and purified by a biotechnology, and finally applied to a clinical diagnostic reagent. The yolk antibody is similar to the IgG of the serum antibody of the mammal, but the yolk antibody obtained by taking the yolk as the raw material has great advantages compared with the antibody derived from the serum of the mammal.
The traditional human diagnostic reagent antibody raw material uses mammal raw material source antibody (sheep, rabbit, etc.), which needs to be obtained after blood serum is separated by blood collection, and has the main problems that: the method needs a culture place with a considerable scale, needs blood sampling, does not accord with the animal welfare principle, and has the defects of detection sensitivity and accuracy interference due to the existence of detection cross factors with mammals belonging to the same genus as human beings.
Disclosure of Invention
The technical problem to be solved by the invention is that the obtained antibody has higher sensitivity than a mammal source antibody, the obtained source is wide and convenient, and the production scale can be flexibly enlarged.
In order to solve the technical problems, the invention provides a preparation method of a yolk antibody for a clinical diagnostic reagent, which comprises the following specific steps:
the first step is as follows: strengthening immunity, immunizing chicken with clinical diagnosis index immunogen according to a specific program, and accumulating specific yolk antibody in the eggs laid by the chicken;
the second step is that: performing titer detection, namely performing agar two-way immunodiffusion titer detection on egg yolk, and collecting qualified egg yolk;
the third step: acidifying, diluting egg yolk with purified water, acidifying, and centrifuging to remove most lipid;
the fourth step: degreasing, namely completely removing residual lipid by using a degreasing technology;
the fifth step: deep clarification and filtration, and taking out fine particulate matters in the degreased material liquid;
and a sixth step: purifying and concentrating by tangential flow ultrafiltration, purifying the yolk antibody, and concentrating the material liquid volume to the level before dilution and acidification of purified water;
the seventh step: and (4) carrying out low-temperature vacuum freeze drying, and carrying out freeze drying through a set program to obtain the yolk antibody freeze-dried powder.
Preferably: the first step of strengthening the immune chicken comprises the following steps: the enhanced immunity is carried out every 7-20 days, the antigen and the adjuvant are mixed in equal volume, 0.5-1.5 ml of breast muscle or infrapteral muscle of the chicken is injected at multiple points, and the dosage of the antigen is 10-1000 ug/egg.
Preferably, the adjuvant in the first step is Freund's complete adjuvant, Freund's incomplete adjuvant, natural source adjuvant, oil emulsion adjuvant, cytokine adjuvant, microbial adjuvant, nucleic acid adjuvant, nano adjuvant, etc.
Preferably, the second step of detecting the egg yolk antibody titer comprises the following steps: adding 0.85% normal saline into egg yolk liquid for gradient dilution, and detecting the titer of the egg yolk to be more than 1:64, collecting eggs, separating egg yolk
Preferably, the third acidification method comprises the following steps: diluting egg yolk with purified water at a ratio of 1: 5-1: 9, adjusting pH to 5.0-6.0, mixing, stirring, standing overnight at 4 ℃, separating supernatant, and taking supernatant as acidified yolk liquid.
Preferably, the fourth degreasing technology is as follows: adding fumed silica with the mass concentration of 1-20 g/l into the acidified yolk liquid, uniformly mixing, centrifuging, collecting supernatant, and taking the supernatant to obtain the completely defatted yolk antibody liquid.
Preferably, the fifth deep clarification filtration step comprises the following steps: filtering the completely defatted egg yolk antibody liquid by a folding filter element or a membrane with the filtering pressure difference of 0-4 bar, and taking the filtrate to obtain the particle-free clarified egg yolk antibody liquid.
Preferably, the sixth step of tangential flow ultrafiltration purification and concentration comprises the following steps: and (3) ultrafiltering and concentrating the clear egg yolk antibody liquid without particles by using a stacked membrane package with the filtration pressure difference of 0-4 bar and the pore diameter of 10-50 kD, taking the concentrated solution as the purified egg yolk antibody liquid, wherein the volume of the purified egg yolk antibody liquid is the volume before the purified water is added in the second step for dilution.
Preferably, the seventh low-temperature vacuum freeze-drying step comprises the following steps: and (3) pre-freezing for 2-4 hours, cooling to-50 to-80 ℃, maintaining for 2-4 hours, then setting the temperature to rise for 4-8 ℃ per hour, and freeze-drying in a vacuumizing state to obtain white egg yolk antibody powder.
By adopting the method, the production scale can be flexibly enlarged, the product has high production capacity, so that the method has high economic benefit, and meanwhile, under the same immunity condition, the difference of the phylogenetic distance ensures that the chicken antibody is better than the mammal antibody in the aspects of resisting highly conserved protein or polypeptide and the like, and has the advantages of meeting the animal interest protection, having economic advantage, having no cross reaction with the rheumatism factor, having no cross reaction with the human anti-mouse antibody, having no activity of a complement system, having no lectin and having no affinity of the protein A, G.
Detailed Description
The technical solution of the present invention is described in full and clearly below by way of example applications.
Example 1
1 Material
1.1 antigens
C-reactive protein (CRP) from SIGMA reagent (cat # C1617).
1.2 immunization-use laying hens
High-yield Roman brown chickens which are 200-day-old commodities are conventionally bred in chicken farms.
1.3 reagents
Freund's complete adjuvant, Freund's incomplete adjuvant were purchased from SIGMA reagent; the hydrochloric acid was analytically pure, fumed silica was purchased from SIGMA reagent, cat # S5130.
1.4 Experimental instruments
The centrifuge is a J6-MI large capacity freezing centrifuge, the filtering device is a Millipore sterilization filter, the ultrafiltration device is a Millipore Pellicon ultrafiltration system, and the freeze dryer is a Virtis desktop Benchtop Pro.
2 method
The first step of strengthening the immune chicken comprises the following steps:
the antigen priming dose is 10 ug/mouse, the strengthening immunity is carried out every 10 days, the antigen and Freund's complete adjuvant (purchased from SIGMA reagent) are mixed in equal volume and are fully emulsified, 0.5 ml of the breast muscle of the chicken is injected in multiple points, the first strengthening immunity is carried out 10 days after the antigen priming, the antigen dose is 15 ug/mouse, the adjuvant is Freund's incomplete adjuvant (purchased from SIGMA reagent) in equal volume and is fully emulsified, 0.5 ml of the breast muscle of the chicken is injected in multiple points, the second strengthening immunity is carried out 10 days after the first strengthening immunity, the antigen dose is 20 ug/mouse, the adjuvant is Freund's incomplete adjuvant (purchased from SIGMA reagent) in equal volume and is fully emulsified, and the breast muscle of the chicken is injected in multiple points is 0.5 ml.
The second step of the egg yolk antibody titer detection method comprises the following steps:
adding 0.85% physiological saline into egg yolk solution for gradient dilution, wherein the dilution is 2 times, 4 times, 8 times, 16 times, 32 times and 64 times, and the egg yolk detection titer exceeds 1: and 64, collecting the eggs, and separating egg yolks of the eggs.
The third step of acidification method is:
adding 9 times of purified water into the egg yolk liquid, mixing uniformly, slowly dropwise adding hydrochloric acid into the mixed liquid, adjusting the pH to 5.0-6.0, fully mixing uniformly, stirring for homogenizing, standing overnight at 4 ℃, adding the egg yolk liquid into a centrifugal tube, placing the centrifugal tube into a centrifugal machine (J6-MI high-capacity refrigerated centrifuge) after balancing, setting the centrifugal machine at 4 ℃, centrifuging for 30 minutes at 10000g, filtering and separating supernatant by using a filter (Millipore sterilization filter), and taking the supernatant as acidified egg yolk liquid.
The fourth step degreasing technology comprises the following steps:
adding 400 m with the mass concentration of 2 g/l and the specific surface area of 200-2Adding fumed silica (purchased from SIGMA reagent, product number S5130) with the proportion of 10-20% into per gram, uniformly mixing, standing for 2-4 hours, adding the acidified egg yolk liquid into a centrifuge tube, placing the centrifuge tube into a J6-MI high-capacity refrigerated centrifuge after balancing, setting the centrifuge at 4 ℃, centrifuging for 30 minutes at 10000g, filtering and separating supernatant by a filter (Millipore sterilization filter), and taking the supernatant as the completely defatted egg yolk antibody liquid.
The fifth step is a deep clarification filtration step method:
sequentially filtering and clarifying with folded bag type filter with pore diameter of 2.0 um, 1.2 um, 0.65 um, 0.45 um and 0.22 um, separating completely defatted yolk antibody solution, and collecting clarified solution as granule-free clarified yolk antibody solution.
The sixth step of tangential flow ultrafiltration purification and concentration comprises the following steps:
ultrafiltering the homogeneous clarified yolk antibody liquid with ultrafiltration membrane system (Millipore Pellicon ultrafiltration system) with ultrafiltration membrane having pore diameter of 30 kD, ultrafiltration pressure of less than 4 bar, and 0.5 mm ultrafiltration membrane, and concentrating the concentrated solution to volume before defatting, wherein the concentrated solution is purified yolk antibody;
the seventh step of low-temperature vacuum freeze drying comprises the following steps:
placing the purified yolk antibody concentrated solution into a freeze dryer (American Virtis desktop benchmark Pro) flat plate, setting temperature gradient to reduce the temperature to-50 ℃ within 2 hours, maintaining for 2 hours, then raising the temperature to 5 ℃ per hour, vacuumizing while freeze-drying, freeze-drying for about 24 hours, finally maintaining for 5 hours at room temperature, taking out the freeze-dried yolk antibody, wherein the freeze-dried yolk antibody is white powder which is easy to dissolve in water, and storing in a refrigerator at 4 ℃.
And (3) carrying out effect detection on the obtained product:
one, two-way immunodiffusion potency assay for agar
And (3) re-dissolving the freeze-dried yolk antibody product with 0.85% physiological saline to the volume before freeze-drying, and performing two-way immunodiffusion according to the conventional agar. The results are shown in Table 1.
TABLE 1 original yolk titer and purified yolk antibody product titer
Table 1 the results show that: the original yolk agar two-way immunodiffusion titer is greater than the collection standard 1:64, the yolk antibody titer is minimum 1:64 after the process purification, the quality standard is met, and further clinical biochemical detection data prove that the yolk antibody titer meets the use requirement, and the result is shown below.
Second, biochemical turbidimetric assay
Taking a freeze-dried egg yolk antibody product, re-dissolving the freeze-dried egg yolk antibody product into the volume before freeze-drying by using 0.85 percent physiological saline, preparing a diagnostic reagent according to a process formula, performing biochemical turbidimetry measurement by using a quality control product and a clinical serum sample, wherein the correlation coefficient r of a quality control standard curve is 0.9991, and comparing the detection of the clinical sample by using a conventional goat antiserum diagnostic reagent and an egg yolk antibody diagnostic reagent synchronously, wherein the data of regression curves of the two reagents are shown in a table 2:
TABLE 2 comparison of yolk antibody diagnostic reagents with sheep antiserum diagnostic reagents
The results in Table 2 show that, compared with the traditional sheep antiserum diagnostic reagent, the egg yolk antibody diagnostic reagent has the clinical use effect which is comparable to that of the sheep antiserum reagent, and the egg yolk antibody is an excellent substitute antibody raw material which can be applied to various clinical biochemical detections.
Although specific embodiments of the present invention have been described above, it will be appreciated by those skilled in the art that these are merely examples and that many variations or modifications may be made to the embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims.
Claims (9)
1. A preparation method of a yolk antibody for a clinical diagnostic reagent is characterized by comprising the following steps: the method comprises the following specific steps:
the first step is as follows: strengthening immunity, immunizing chicken with clinical diagnosis index immunogen according to a specific program, and accumulating specific yolk antibody in the eggs laid by the chicken;
the second step is that: performing titer detection, namely performing agar two-way immunodiffusion titer detection on egg yolk, and collecting qualified egg yolk;
the third step: acidifying, diluting egg yolk with purified water, acidifying, and centrifuging to remove most lipid;
the fourth step: degreasing, namely completely removing residual lipid by using a degreasing technology;
the fifth step: deep clarification and filtration, and taking out fine particulate matters in the degreased material liquid;
and a sixth step: purifying and concentrating by tangential flow ultrafiltration, purifying the yolk antibody, and concentrating the material liquid volume to the level before dilution and acidification of purified water;
the seventh step: and (4) carrying out low-temperature vacuum freeze drying, and carrying out freeze drying through a set program to obtain the yolk antibody freeze-dried powder.
2. The method for preparing a yolk antibody for clinical diagnostic reagents according to claim 1, wherein the method comprises the steps of: the strengthening method of the first step of immunizing the chicken comprises the following steps: the enhanced immunity is carried out every 7-20 days, the antigen and the adjuvant are mixed in equal volume, 0.5-1.5 ml of breast muscle or infrapteral muscle of the chicken is injected at multiple points, and the dosage of the antigen is 10-1000 ug/egg.
3. The method for preparing a yolk antibody for clinical diagnostic reagents according to claim 2, wherein: the adjuvant in the first step is Freund's complete adjuvant, Freund's incomplete adjuvant, natural source adjuvant, oil emulsion adjuvant, cytokine adjuvant, microbial adjuvant, nucleic acid adjuvant, nano adjuvant, etc.
4. The method for preparing a yolk antibody for clinical diagnostic reagents according to claim 1, wherein the method comprises the steps of: the second step of detecting the egg yolk antibody titer comprises the following steps: adding 0.85% normal saline into egg yolk liquid for gradient dilution, and detecting the titer of the egg yolk to be more than 1: and 64, collecting the eggs, and separating egg yolks of the eggs.
5. The method for preparing a yolk antibody for clinical diagnostic reagents according to claim 1, wherein the method comprises the steps of: the third acidification method comprises the following steps: diluting egg yolk with purified water at a ratio of 1: 5-1: 9, adjusting pH to 5.0-6.0, uniformly mixing and stirring, standing overnight at 4 ℃, separating supernatant, and taking supernatant as acidified yolk liquid.
6. The method for preparing a yolk antibody for clinical diagnostic reagents according to claim 1, wherein the method comprises the steps of: the fourth degreasing technology comprises the following steps: adding fumed silica with the mass concentration of 1-20 g/l into the acidified yolk liquid, uniformly mixing, centrifuging, collecting supernatant, and taking the supernatant to obtain the completely defatted yolk antibody liquid.
7. The method for preparing a yolk antibody for clinical diagnostic reagents according to claim 1, wherein the method comprises the steps of: the fifth step of deep clarification and filtration comprises the following steps: filtering the completely defatted egg yolk antibody liquid by a folding filter element or a membrane with the filtering pressure difference of 0-4 bar, and taking the filtrate to obtain the particle-free clarified egg yolk antibody liquid.
8. The method for preparing a yolk antibody for clinical diagnostic reagents according to claim 1, wherein the method comprises the steps of: the sixth step of tangential flow ultrafiltration purification and concentration comprises the following steps: and (3) ultrafiltering and concentrating the clear egg yolk antibody liquid without particles by using a stacked membrane package with the filtration pressure difference of 0-4 bar and the pore diameter of 10-50 kD, taking the concentrated solution as the purified egg yolk antibody liquid, wherein the volume of the purified egg yolk antibody liquid is the volume before the purified water is added in the second step for dilution.
9. The method for preparing a yolk antibody for clinical diagnostic reagents according to claim 1, wherein the method comprises the steps of: the seventh step of low-temperature vacuum freeze drying comprises the following steps: and (3) pre-freezing for 2-4 hours, cooling to-50 to-80 ℃, maintaining for 2-4 hours, then setting the temperature to rise for 4-8 ℃ per hour, and freeze-drying in a vacuumizing state to obtain white egg yolk antibody powder.
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| CN112485455A (en) * | 2020-12-09 | 2021-03-12 | 深圳市亚辉龙生物科技股份有限公司 | New coronavirus antibody quality control product and preparation method thereof |
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| 陈亮等: "卵黄抗体纯化和浓缩的研究进展", 《黑龙江畜牧兽医》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112485455A (en) * | 2020-12-09 | 2021-03-12 | 深圳市亚辉龙生物科技股份有限公司 | New coronavirus antibody quality control product and preparation method thereof |
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