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Aminoacid mimic epitope of human B lymphocyte stimulating factor receptor and use thereof

A technology of B lymphocytes and stimulators, which is applied in the fields of biopharmaceuticals and genetic engineering, and can solve the problems of insufficient display efficiency

Inactive Publication Date: 2009-01-21
JIANGSU PROVINCE INST OF TRADITIONAL CHINESE MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In addition, Hufton et al published a research report on the display system of P6 display system, but the display efficiency of this system is not as good as that of P3 and P8

Method used

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  • Aminoacid mimic epitope of human B lymphocyte stimulating factor receptor and use thereof
  • Aminoacid mimic epitope of human B lymphocyte stimulating factor receptor and use thereof
  • Aminoacid mimic epitope of human B lymphocyte stimulating factor receptor and use thereof

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Embodiment

[0040] To realize the antibody purification, peptide library screening, animal immunization and antibody identification of the present invention, it is completed according to the following steps:

[0041] 1. Purification and identification of BAFF-R antibody:

[0042] After BAFF-R monoclonal antibody ascites was diluted 1:10 with loading buffer, it was applied to MONO Q ion exchange column for anion exchange chromatography, followed by loading, binding and elution. Gradient elution was performed with 50 mmol Tris-HCl buffer (pH 8.0) containing 0.35 mol NaCl, and 3 peaks were obtained. The eluate was subjected to 12% SDS-PAGE ( figure 1 ), the results confirmed that: the third peak contained BAFF-R antibody with higher purity, and the purity of BAFF-R antibody obtained was about 90% after scanning by Kodak imaging system.

[0043] 2. Biopanning of Ph.D-7 peptide library

[0044] Phage Random Heptapeptide Library (Ph.D.-7 TM phage display peptide library): purchased from Ne...

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Abstract

The invention relates to a mimic eptitope peptide of BAFF-R and a DNA coding the peptide and the application of the mimic eptitope peptide in the preparation of antitumor bacterins and drugs. The mimic eptitope of 7 amino acids of BAF-R is a mimic eptitope of molecule BAFF-R with high affinity with a monoclonal antibody of BAFF-R, and is obtained through selection from a phage random display 7-peptide bank with a monoclonal antibody of BAFF-R as the antigen, wherein the sequence of the amino acids is Gly Tyr Thr Arg Trp Gly Cys. The 7-amino-acid mimic eptitope can also be used to construct a poly-peptide vaccine. The clonal inhibition rates of the phage display peptide provided by the invention are all more than 50 percent, and can specifically inhibit the combination between antibody and antigen for a larger extent, greatly improve the proliferation of mouse spleen lymphocytes without adjuvant, the mimic antigen is good in immunogenicity, as illustrated by the induced cell immune response after mice are vaccinated with the mimic antigen.

Description

technical field [0001] The invention relates to the fields of biopharmaceuticals and genetic engineering, in particular to a mimetic epitope peptide of B lymphocyte stimulating factor receptor (BAFF-R) protein, its coding DNA and its application in preparing anti-tumor vaccines and medicines. Background technique [0002] 1. Research on BAFF-R molecules and their inhibitors [0003] Human BAFF-R is a type III membrane protein with a full length of 184 amino acid residues, and its homology to human and mouse is 56%. The CRD region of its extracellular part has only 4 cysteines, which is the least among known TNFR superfamily members. BAFF-R gene is located in human 22q13.1-13.31, expressed in spleen, lymph nodes and peripheral blood, but not detected in bone marrow and fetal liver. Different from the other two receptors of B lymphocyte stimulating factor, TACI and BCMA, BAFF-R only binds to BAFF with high affinity, and it plays an important role in the development and funct...

Claims

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Application Information

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IPC IPC(8): C07K7/06A61K39/00A61K38/08A61P35/00
Inventor 曹鹏夏志南
Owner JIANGSU PROVINCE INST OF TRADITIONAL CHINESE MEDICINE
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