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Preparation method of large capacity serum antibody

A serum antibody and large-capacity technology, which is applied in the field of large-capacity serum antibody preparation, can solve the problems of not being able to meet the large-capacity serum antibody, affecting the quality of the extracted serum antibody, and incomplete removal of fibrin, etc. The method is simple and the removal is thorough , good stability

Inactive Publication Date: 2017-10-27
SOUTHWEST UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, the most common methods for extracting and preparing serum antibodies are freeze-thaw method, cold ethanol method, and natural precipitation method. The first two methods are low in cost, but they do not completely remove fibrin in plasma, and it is easy to appear in a short time after preparation. The precipitation of flocs affects the quality of the extracted serum antibody and subsequent related research; the natural precipitation method has the least impact on the total protein amount of the extracted serum, and the quality of the precipitated serum antibody is very good, but the amount of the extracted serum antibody is the least. Lost more, can not meet the needs of the preparation of large-capacity serum antibodies

Method used

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  • Preparation method of large capacity serum antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] The preparation method (thrombin method) of large-capacity serum antibody, concrete steps are as follows:

[0029] (1) Take 5, 2, 3, 4 parts / head or 2, 2, 2, 4 heads / parts of normal healthy fattened pigs to be immunized with swine fever vaccine (note: the vaccine is live vaccine of swine fever from rabbit origin , specifications; 10 doses / bottle, ≥0.02g tissue poison / head dose, the vaccine production unit is: Shanghai Haili Biotechnology Co., Ltd.), immunize once a week, and immunize with 2-3 times the dose of the first immunization in the fifth week Swine fever vaccine, after 3 days of double immunization, slaughter and collect fresh pig blood; before slaughter, rinse the whole body of the fat pig to be slaughtered with clean tap water, and use Sur to disinfect; bloodletting knives or other slaughtering supplies are disinfected with alcohol or other disinfectants;

[0030] (2) Add the fresh pig blood collected in step (1) into 3.8% sodium citrate in a volume ratio of 1...

Embodiment 2

[0050] In order to determine the best conditions for the preparation of serum antibodies by the thrombin method, optimize the dosage and storage temperature of thrombin:

[0051] 1) Optimization of the optimal dosage of thrombin

[0052] Take 10 500mL sterilized beakers, add 150mL of spare plasma respectively, add 1500IU, 2000IU, 2500IU, 3000IU, 3500IU, 4000IU, 4500IU, 5000IU, 5500IU, 6000IU thrombin into the plasma respectively, stir and mix evenly, 37 Stand still at ℃ for 0.5-1h, then filter with filter paper with a pore size of 0.88μm for 15-24h, collect the filtrate that has been completely filtered, transfer it into a 250mL reagent bottle with the corresponding number, and observe it at 4°C for 12h, 24h, 48h when collecting. Record the test results. The results show that when thrombin and plasma are used at a ratio of less than 30IU: 1mL, there are different degrees of precipitates in the collected filtrate after the precipitation is complete and standing at 4°C for 48h-...

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Abstract

The invention discloses a method for preparing a large-capacity serum antibody, which comprises the following steps: first collecting immunized fresh blood, collecting the plasma, and freezing and storing it for later use; then filtering the collected plasma, and collecting the filtrate to obtain a primary filtered serum product; Add thrombin to the prepared primary filtered serum product, stir evenly, and then apply it warmly for 0.5 to 4 hours at 20 to 37°C, then filter with filter paper with a pore size of 0.22 to 0.88 μm, collect the filtrate, and obtain serum antibodies; using the present invention The serum antibody prepared by the method has large quantity, good quality and good stability, is suitable for the preparation of large-capacity serum antibody, and has a good market prospect.

Description

technical field [0001] The invention belongs to the field of biology, and in particular relates to a method for preparing large-capacity serum antibodies. Background technique [0002] Serum antibodies are the most primary products in antibody drugs, and are only prepared on a small scale in laboratories for scientific research. Fibrinogen is one of the main components that distinguish plasma from serum. It is a hexamer with a molecular weight of 340 kDa formed by a, β and γ chains connected by 29 pairs of disulfide bonds. It acts as a receptor for platelet membrane glycoprotein Ⅱb / Ⅲa. The body participates in the activation and aggregation of platelets and the third stage of coagulation under the action of thrombin and other components of the animal body. [0003] With the rapid development of the intensification of the breeding industry, immune failure, sporadic outbreaks of major animal diseases, the prohibition of antiviral drugs in veterinary clinics, and mixed infecti...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/06
Inventor 魏光河
Owner SOUTHWEST UNIV
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