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Human IgA immunoglobulin combination molecule having intramolecular affinity effect

A technology of immunoglobulin and binding molecule, applied in the field of human IgA immunoglobulin binding molecule and its preparation

Inactive Publication Date: 2011-07-06
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Based on this, further obtaining high-affinity new hIgA affibody combination molecules to substantially improve the binding activity with hIgA is a problem worthy of further research, and there have been no reports at home and abroad.

Method used

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  • Human IgA immunoglobulin combination molecule having intramolecular affinity effect
  • Human IgA immunoglobulin combination molecule having intramolecular affinity effect
  • Human IgA immunoglobulin combination molecule having intramolecular affinity effect

Examples

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Embodiment 1

[0169] Application of phage molecular evolution to obtain human IgA avidosome repeats with intramolecular affinity effect

[0170] 1. Gene synthesis of the SEG1 and SEG2 fragments of hIgA affibodies According to the hIgA affibodies provided by the literature [Ronnmark J, Gronlund H, Uhlen M, Nygren PA. (2002) Eur J Biochem 269(11): 2647-2655.] The amino acid sequence of the hIgA affinity body SEG1 and SEG2 genes (both containing 174 nucleotides) of the E. coli preferred codon sequence was designed, and was entrusted to Shanghai Yingjun Biotechnology Co., Ltd. to synthesize.

[0171] 2. Construction of recombinant pMD-18T cloning vector The synthesized SEG1 and SEG2 gene sequences were cloned into pMD-18T vector, named pMD-18T / SEG1 and pMD-18T / SEG2. For the construction process of the recombinant pMD-18T cloning vector, see figure 1 . Three recombinant plasmids of pMD-18T / SEG1 and pMD-18T / SEG2 were picked, and entrusted to Shanghai Yingjun Biotechnology Co., Ltd. for sequence...

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Abstract

The invention discloses a human IgA immunoglobulin combination molecule having intramolecular affinity effect, a preparation method and an application thereof. The invention also discloses genes coding the human IgA immunoglobulin combination molecule, a preparation method and an application of the human IgA immunoglobulin combination molecule based on bacteriophage molecule evolution. The human IgA immunoglobulin combination molecule of the invention, especially repeated molecule of human IgA affibody, has intramolecular affinity effect when binding the human IgA, demonstrates very high human IgA binding activity, can be used for purification of high-specific IgA and research and development of detecting reagent, and purification for detection of human IgA antibody by ELISA adsorption method, immunity chromatography, immunohistochemical method and the like.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and relates to a human IgA immunoglobulin binding molecule with intramolecular affinity effect and a preparation method and application thereof. Background technique [0002] IgA is the main immunoglobulin (immunoglobulin, Ig) in the human mucosal surface and secretion fluid. It not only plays an important role in the body's immune defense, but is also closely related to the diagnosis and prognosis of many diseases. Therefore, research and development of highly efficient and specific human IgA (hIgA) purification and detection reagents have important application value. Staphylococcus aureus protein A (SPA) is one of the bacterial surface proteins with Ig binding activity. According to literature reports, a new protein A single domain (named Z domain) containing 58 amino acids was synthesized using the amino acids with the highest frequency in each position of the five Ig-binding single domai...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/47C07K1/14C12N15/12C12N15/63C12N15/70C12N15/74C12N1/21C40B30/04C12Q1/68G01N33/53
Inventor 潘卫曹洁温宗梅陈秋莉蒋少华王锦红戚中田
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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