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Anti-human coagulation factor VII monoclonal antibody, preparation method thereof and use thereof

A monoclonal antibody and human coagulation factor technology, applied in the field of genetic engineering, can solve the problems of unsatisfactory antibodies and lack of specific human coagulation factor VII, etc.

Inactive Publication Date: 2010-12-08
SUZHOU ZELGEN BIOPHARML
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the existing antibodies are still unsatisfactory, so there is still a lack of effective methods for detecting human coagulation factor VII with strong specificity

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1: Preparation of Monoclonal Antibody

[0044] (1) Immunized animals:

[0045] On the first day, antigen 50 μg / mouse, 0.25ml antigen + 0.25ml complete Freund’s adjuvant / mouse, mixer at 6000 rpm for 10 minutes, fully mixed, subcutaneous multi-point injection, 5 mice / antigen; day 28 , antigen 50μg / mouse, 0.25ml antigen + 0.25ml incomplete Freund's adjuvant / mouse, stirrer 6000 rpm for 10 minutes, fully mixed, subcutaneous multi-point injection, 5 mice / antigen; on the 60th day, antigen 50 μg / mouse, 0.25ml antigen + 0.25ml incomplete Freund’s adjuvant / mouse, stirrer 6000 rpm for 10 minutes, fully mixed, multi-point subcutaneous injection, 5 mice / antigen; on the 90th day, antigen 50μg / One mouse, 0.25ml antigen + 0.25ml incomplete Freund's adjuvant per mouse, mixer at 6000 rpm for 10 minutes, fully mixed, multi-point subcutaneous injection, 5 mice per antigen. Four days before fusion, 30 μg of antigen was injected intraperitoneally.

[0046] (2) Cell fusion:

[00...

Embodiment 2

[0106] Example 2: Purification and detection of antibodies

[0107] Ascitic fluid was prepared from 6 hybridoma cells, purified with commercial Protein A, and the operation method was referred to its instruction manual, and the purified monoclonal antibody was obtained as a result.

[0108] Binding specificity and affinity tests were carried out after the antibodies were purified, and the results showed that the specificity and affinity of the six mAbs were all very good, especially the monoclonal antibody F7-2 had the best specificity and affinity, which was better than other general monoclonal antibodies (such as hybrid The monoclonal antibody produced by tumor cells D) was about an order of magnitude higher.

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PUM

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Abstract

The invention provides an anti-human coagulation factor VII monoclonal antibody, a preparation method thereof and use thereof. The monoclonal antibody has a characteristic of high human coagulation factor VII antigen binding specificity. The invention also provides a corresponding monoclonal antibody fragment and an immunoconjugate. The invention also provides a detection kit for detecting the human coagulation factor VII.

Description

technical field [0001] The invention relates to the field of genetic engineering. More specifically, the present invention relates to an anti-human blood coagulation factor VII monoclonal antibody and its preparation method and use. Background technique [0002] Coagulation of wounds is initiated after the formation of a complex between human coagulation factor VII and tissue factor. This complex activates factor X to Xa and factor IX to IXa. Xa and other factors form a complex that converts prothrombin to thrombin, which induces local hemostasis by ultimately converting fibrinogen to fibrin to form a fibrin clot [Robert H, et al, Molecular biology and biochemistry of the coagulation factors and pathways of haemostasis. William's Hematology, 2001: 1409-1434]. Human factor VII can be used in patients with congenital hemophilia who have inhibitors of factor VIII or IX > 5 BU, patients with congenital hemophilia who are expected to respond to injections of factor VIII or ...

Claims

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Application Information

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IPC IPC(8): C07K16/36C07K19/00C12N5/20G01N33/577
Inventor 盛泽林
Owner SUZHOU ZELGEN BIOPHARML
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