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A kind of monoclonal antibody against human btf and its application

A monoclonal antibody, species-specific technology, applied in the field of antibody engineering and medicine, can solve problems such as poor stability and chromosome deletion

Inactive Publication Date: 2015-08-19
BEIJING INST OF GENOMICS CHINESE ACAD OF SCI CHINA NAT CENT FOR BIOINFORMATION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] BTF was newly discovered by KASOF and others when they used the yeast two-hybrid system to search for the interacting protein of the virus anti-apoptotic protein E1B19K. It is located in the segment of chromosome 6q21-23. This segment has poor stability and is prone to chromosome deletions in various tumors.

Method used

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  • A kind of monoclonal antibody against human btf and its application
  • A kind of monoclonal antibody against human btf and its application
  • A kind of monoclonal antibody against human btf and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Example 1 Preparation and Screening of Mouse Monoclonal Antibody

[0017] (1) Antigen preparation

[0018] The human BTF (Bcl-2-associated transcription factor 1) gene was amplified from the human gene transcriptome by PCR. The upstream primer was: 5'-GGCCGCTAGCATGGACTACAAGGACGACGATGACAAGGGTCGCTCCAATTCTAG-3'; the downstream primer was: 5'-GGCCGGATCCTTATTCCTTTTCTTCCTTGC-3'. Using NheⅠ and BamHI double enzyme digestion, the gene fragment was connected into the eukaryotic expression vector pIRESneo3, PCR and double enzyme digestion identified positive clones and then sequenced. After the sequence was determined correctly, the plasmid was extracted, and Lipofectamine2000 (Invitrogen) mediated transfection into 293T cells, G418 Screen stable clones and transfer to culture flasks for expression. Cells were collected by centrifugation, non-denatured cells were lysed, centrifuged at 10,000rpm for 10 minutes, BTF was extracted by FLAG affinity chromatography, the eluate was use...

Embodiment 2W

[0028] Embodiment 2Western-Blot detects the expression of BTF

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PUM

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Abstract

The invention relates to the field of antibody engineering and medicines, and specifically provides a specific monoclonal antibody of anti-human BTF (Bcl-2-associated transcription factor 1) and associated hybridoma thereof. The invention further provides application of the monoclonal antibody in preparing reagents for immunological detection. The monoclonal antibody provided by the invention can be used for specifically identifying human BTF.

Description

technical field [0001] The invention relates to the fields of antibody engineering and medicine. More specifically, the present invention relates to obtaining a mouse monoclonal antibody that recognizes an epitope of human BTF (Bcl-2-associated transcription factor 1). Background technique [0002] BTF was newly discovered by KASOF and others when they used the yeast two-hybrid system to search for the interacting protein of the virus anti-apoptotic protein E1B19K. It is located in the segment of chromosome 6q21-23. This segment has poor stability and is prone to chromosome deletions in various tumors. . BTF has two transcripts of 5kb BTFL and 3kb BTFS; BTFL encodes 918 amino acids with a molecular weight of 106KD; BTFS encodes a protein with a molecular weight of 101KD and its carboxy-terminal 797-846 amino acids are missing. BTFL is strongly expressed in various epithelial tissues, such as skeletal muscle, placenta, heart, brain, lung, kidney and pancreas, while BTFS is ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/18C12N5/20G01N33/68G01N33/577
Inventor 王绪敏绍长君任鲁风刘贵明王延强
Owner BEIJING INST OF GENOMICS CHINESE ACAD OF SCI CHINA NAT CENT FOR BIOINFORMATION