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N-end monoclonal antibody MA7B1 of anti-human multicapsular protein-1

A technology of monoclonal antibody and polycystic protein, which is applied in the direction of anti-animal/human immunoglobulin, measuring devices, instruments, etc., can solve the problems of poor experimental stability and achieve good stability, strong specificity, and high sensitivity Effect

Inactive Publication Date: 2004-09-08
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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Problems solved by technology

However, most antibodies can only meet the detection of one or two of the comprehensive performance indicators such as immunohistochemistry, enzyme-linked immunosorbent assay (ELISA), Western blot (Western blot) and immunoprecipitation (IP), but can be applied Antibodies based on IP are even rarer and less stable in experiments

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  • N-end monoclonal antibody MA7B1 of anti-human multicapsular protein-1
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  • N-end monoclonal antibody MA7B1 of anti-human multicapsular protein-1

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Embodiment Construction

[0035] The preparation of anti-human PC-1 monoclonal antibody MA7B1 is now described in detail.

[0036] 1. Materials

[0037] (1) Plasmids and bacterial strains: fusion protein expression vector pQE30, the vector contains PT5 promoter, 2 lac induction sequences, nucleotide sequence encoding RBS, initiation codon ATG, encoding 6 consecutive histidines from upstream to downstream Nucleotide sequence of acid, multiple cloning site MCS and stop codon. This plasmid also contains the ampicillin resistance marker gene and the replicon Col E1. The expression host strain M15 contains the repressor plasmid pREP4, which contains the kanamycin resistance marker gene. PQE30 and M15 were purchased from Qiagen, Germany.

[0038] (2) Animals: female New Zealand white rabbits were purchased from the Experimental Animal Center of Shanghai Second Medical University.

[0039](3) Main reagents: total RNA extraction kit and gel recovery kit are from Huashun Company, and One-Step RT-PCR kit and...

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Abstract

The present invention relates to medicine molecular biology and immulogical detection technology, and is one kind of 1N-end monoclonal antibody MA7B1 of anti-human polycystic protein-1. According to cDNA sequence of human polycystic kidney disease-1 gene and the molecular structure of its encoded product, polycystic protein-1 (PC-1), the gene is cloned to cDNA encoding PC-1 and its prokaryotic expression carrier is constituted by means of molecular biology technology. N-end fusion protein of PC-1 is induced in colibacillus and anti-PC-1 N-end monoclonal antibody MA7B1 is prepared by means of hybrid tumor technology. Comprehensive performance test shows that the antibody has stable performance, high sensitivity and powerful specificity in immunohistochemistry, enzyme-linked immunosorbent assay, immunoblotting and immunoprecipiation experiments, so that it may be used in preparing reagent for polycystic kidney disease mechanism research.

Description

technical field [0001] The invention relates to the technical fields of medical molecular biology and immunological detection, is a kind of anti-human PC-1 N-terminal monoclonal antibody MA7B1, and can be used for preparing reagents for studying the pathogenesis of autosomal dominant hereditary polycystic kidney disease. Background technique [0002] Autosomal dominant polycystic kidney disease (ADPKD) is a hereditary kidney disease that seriously endangers human health. Patients with bilateral kidneys are densely covered with cysts, and often develop into end-stage renal failure in late middle age. Due to its pathogenic The gene is widely expressed in the body, and often involves other epithelial organs, including liver, pancreas, ovary and choroid plexus, etc., causing great harm. At present, it has been clear that the abnormal function of polycystin and the resulting abnormality of a series of biological signal transduction are the key links ...

Claims

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Application Information

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IPC IPC(8): C07K16/18G01N33/577
Inventor 梅长林赵海丹
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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