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Antibody heavy chain, light chain variable region and antibody for assisting pbc diagnosis

A variable region and heavy chain technology, applied in the field of biomedicine, can solve problems such as poor therapeutic effect of immunosuppressants

Active Publication Date: 2014-10-08
FOURTH MILITARY MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although PBC is an autoimmune disease, the effect of immunosuppressant therapy is not good

Method used

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  • Antibody heavy chain, light chain variable region and antibody for assisting pbc diagnosis
  • Antibody heavy chain, light chain variable region and antibody for assisting pbc diagnosis
  • Antibody heavy chain, light chain variable region and antibody for assisting pbc diagnosis

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Embodiment Construction

[0056] The present invention will be described in detail below, which is an explanation of the present invention rather than a limitation. The present invention is specifically implemented according to the following steps:

[0057] 1. Preparation of monoclonal antibodies

[0058] 1.1 Separation and preparation of liver membrane proteins from PBC patients by sucrose density gradient centrifugation.

[0059] 1.2 BALB / C mice (6-8 weeks old, female) were immunized with isolated PBC patient liver membrane protein as immunogen.

[0060] First immunization: Mix 100 μg of immunogen with an equal volume of Freund’s complete adjuvant (purchased from Sigma, USA), and inject BALB / C mice subcutaneously at multiple points;

[0061] The second immunization: After 3 weeks, take 50 μg of immunogen and mix it with an equal volume of Freund’s incomplete adjuvant (purchased from Sigma, USA), and then inject BALB / C mice subcutaneously at multiple points;

[0062] The third immunization: 3 weeks...

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Abstract

The invention relates to the technical field of biomedicine, in particular to a monoclonal antibody for diagnosing PBC (Primary Biliary Cirrhosis). The DNA sequences of three complementary determining regions (CDR) for the variable region of the light chain of the antibody for PBC diagnosis provided by the invention are respectively as follows: a a g g c c a g t c a g a a t g t g t a t a c t g c t g t a g c c; t c g g c a t c c a a t c g g t a c a c t; and c a g c a a t a t a g c a g c t a t c c a t t c a c g. The DNA sequences of three complementary determining regions (CDR) for the variable region of the heavy chain are respectively as follows: g g c t t c a a c a t t a g a g a c a c c t a t a t a c a c; a g g a t t g a t c c t g c g g t t g t t a a t a c t a a a t a t g a c c c g a a t t t c c a g g g c; and t a c t t t g a c t a c. Experiment proves that the antibody provided by the invention can effectively assist the diagnosis of PBC, and when antigen shows strong positive expression (+++), a prompt that PBC is possible is provided, and then further diagnosis is needed.

Description

【Technical field】 [0001] The invention relates to the technical field of biomedicine, in particular to a monoclonal antibody used for auxiliary diagnosis of PBC. 【Background technique】 [0002] Primary biliary cirrhosis (PBC) is a chronic intrahepatic cholestatic liver disease mediated by an autoimmune response. The course of the disease is progressive, and can eventually develop into liver fibrosis and cirrhosis. Studies have shown that the detection rate of PBC is on the rise, up to 940 / 1 million population, and the death caused by PBC accounts for 2% of the deaths from liver cirrhosis. Therefore, as one of the important non-viral liver diseases, PBC's harm cannot be ignored. However, due to the diversity of PBC manifestations and the lack of specific serological, pathological, and clinical changes, the current diagnosis of PBC still adopts the principle of comprehensive judgment and uses the method of exclusion to confirm the diagnosis, which greatly limits its clinical ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/13C07K16/18G01N33/68G01N33/577
Inventor 韩英王敬博时永全王璐周新民王学昌
Owner FOURTH MILITARY MEDICAL UNIVERSITY