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Method for preparing antihuman lymphocyte antibodies

A technology of lymphocytes and antibodies, applied in the field of preparation of anti-human lymphocyte antibodies, to achieve the effects of saving production costs, reducing product side effects, and reducing antibody titers

Inactive Publication Date: 2009-11-04
王太林
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The object of the present invention is a preparation method of anti-human lymphocyte antibody, which overcomes the deficiency that the antibody produced by the existing method contains a large amount of mixed antibodies such as red blood cell antibody and platelet antibody

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0018] Rabbit-anti-human lymphocyte antibody preparation: Collect human peripheral blood T lymphocytes and pass them through a nylon wool column, add 100 U of cytokine IL-1 with Freund's complete adjuvant, mix and stir with the cell suspension at a ratio of 1:1, The total dose of immunization is 5ml, and the subcutaneous injection is 3×10 7 cells to complete basic immunity. After 2 weeks, add 100U of cytokine IL-1 with Freund's incomplete adjuvant, mix and stir with the cell suspension at a ratio of 1:1, and the total immune dose is 5ml, and subcutaneously inject 3×10 7 cells to complete booster immunization. After 10 days, use 20ml Hanks solution to make HH lymphoblast suspension, and inject 5×10 cells directly into the ear vein. 7 cells to complete booster immunization again. Blood was collected 10 days later, and the antibody quality was tested according to the test method stipulated in the 2005 edition of "The Pharmacopoeia of the People's Republic of China". The test ...

Embodiment 2

[0025] Pig-anti-human lymphocyte antibody preparation: Collect human peripheral blood T lymphocytes and pass them through nylon hair column, add 100 U of cytokine IL-1 with Freund's complete adjuvant, mix and stir with the cell suspension at a ratio of 1:1, The total dose of immunization is 20ml, subcutaneous injection 15×10 8 cells to complete basic immunity. After 2 weeks, add 100 U of cytokine IL-1 with Freund's incomplete adjuvant, mix and stir with the cell suspension at a ratio of 1:1, and inject a total dose of 20 ml for subcutaneous injection of 15×10 8 cells to complete booster immunization. After 10 days, use 20ml Hanks solution to prepare HH lymphoblastoid cell suspension, and inject 25×10 8 cells to complete booster immunization again. Blood was collected 10 days later, and the antibody quality was tested according to the test method stipulated in the 2005 edition of "The Pharmacopoeia of the People's Republic of China". The test results are as follows:

[002...

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Abstract

The invention provides a method for preparing antihuman lymphocyte antibodies. The method comprises the following: 1) a step of performing basic immunization, which is to adopt purified human peripheral blood T lymphocytes or HH lymphoblast for subcutaneous injection; 2) a step of strengthening immunization, which is to adopt the HH lymphoblast for the subcutaneous injection after two weeks; 3) a step of strengthening immunization again, which is to adopt the HH lymphoblast for auricular intravenous injection after ten days; and 4) a step of collecting plasma, which is to sample blood for detection after ten days, take blood after titer meets requirements and collect the plasma on standby. The method adopts the high-purity lymphocytes as antigens, can significantly reduce the titer of erythrocyte antibodies and platelet antibodies in the plasma, leaves out a hybrid antibody absorption step during the preparation of finished products, and reduces the side effects of the products while significantly reducing production cost.

Description

technical field [0001] The invention belongs to the technical field of raw material preparation of biological preparations, and in particular relates to a preparation method of an anti-human lymphocyte antibody. Background technique [0002] Anti-lymphocyte immunoglobulin (ATG) is a kind of biological immunosuppressant, which can activate complement after combining with T lymphocytes of patients, inhibit or remove T lymphocytes in human body, and is mainly used clinically for rejection of tissue and organ transplantation , acute aplastic anemia, autoimmune hemolytic anemia, idiopathic thrombocytopenic purpura and other malignant immune diseases. [0003] At present, the domestic ATG preparation method is as follows: use human thymus lymphocytes as the immunogen, immunize pigs or rabbits to obtain the raw material serum of anti-human lymphocyte immunoglobulin products, and the raw materials become pig anti-human lymphocytes after purification and refining. Immunoglobulin and...

Claims

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Application Information

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IPC IPC(8): C07K16/18C07K16/06
Inventor 王太林
Owner 王太林
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