Human-derived anti-amylin antibody and preparation method thereof

An amylin, human-derived technology, applied in the field of immunoglobulin, can solve the problems of expensive amylin kits, poor antibody stability and specificity, and human-derived amylin antibodies are prone to cause allergic reactions, etc. Good antigen specificity and antigen binding activity, the effect of broad application prospects

Inactive Publication Date: 2017-01-04
GENERAL HOSPITAL OF TIANJIN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The present invention aims to solve the problems that the existing amylin kits are relatively expensive, and the human-derived amylin antibody prepared by the traditional method is likely to cause allergic reactions, and the stability and specificity of the antibody are poor. A human-derived amylin antibody is proposed. Anti-amyloid antibody and preparation method thereof

Method used

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  • Human-derived anti-amylin antibody and preparation method thereof
  • Human-derived anti-amylin antibody and preparation method thereof
  • Human-derived anti-amylin antibody and preparation method thereof

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Experimental program
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Embodiment Construction

[0025] 1. Experimental materials

[0026] Phagemid vector pComb3XSS (4900bp) was purchased from Scripps Research Institute, USA; Escherichia coli XL1-Blue and helper phage VCSM13 were purchased from Stratagene, USA; TRNzol-A + Total RNA Reagent, Quantscript RT Kit, 2×Taq PCR MasterMix, common agarose gel recovery kit, and high-purity plasmid mini-prep kit were all purchased from TIANGEN; restriction enzymes SacⅠ, XbaⅠ, XhoⅠ, SpeⅠ All were purchased from TaKaRa Company; Amylin fragment (8-37) was purchased from Sigma Company; HA-Tag Mouse mAb (26D11) was purchased from Abmart Company; HRP-goat anti-mouse IgG (H+L) was purchased from Beijing Dingguo Company; HRP-labeled goat anti-human Fab IgG antibody was purchased from Sigma; other reagents were of domestic analytical grade.

[0027] 2. Experimental method

[0028] 1. PCR amplification of human IgG Fab segment gene:

[0029] Lymphocytes were separated from the peripheral blood of normal healthy people with lymphocyte separ...

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Abstract

The invention discloses a human-derived anti-amylin antibody and a preparation method thereof, and belongs to the field of immune globulin. The heavy chain variable region amino acid sequence of the antibody is shown as SEQ ID NO:1, the light chain variable region amino acid sequence of the antibody is shown as SEQ ID NO:2, the heavy chain variable region DNA sequence of the antibody is shown as SEQ ID NO:3, and the light chain variable region DNA sequence of the antibody is shown as SEQ ID NO:4. The heavy chain gene is an IgG1 subclass VH3 gene family, and the light chain gene is a Lambda gene family. The human-derived amylin antibody is screened out by establishing a high-capacity phage antibacterial library, the antibody has good antigen specificity and antigen combination activity, production of an amylin detecting kit is guaranteed, and wide application prospects are achieved.

Description

technical field [0001] The invention belongs to the field of immunoglobulins, in particular to a human-derived anti-amylin antibody and a preparation method thereof. Background technique [0002] Amylin is a hormone secreted by pancreatic islet B cells. It is stored together with insulin in the secretory granules of B cells. The two are released to the outside of pancreatic islet B cells in a certain proportion under the stimulation of glucose and non-glucose hormones. The metabolic process in the body is similar to C-peptide. Studies have found that the metabolism of amylin in the liver is much slower than that of insulin, and amylin is mainly excreted through the kidneys. It has been found that amylin is a pleiotropic hormone that participates in the body's material and energy metabolism with multiple roles, but its exact biological function is still unclear. The main component of islet amyloid (IA) is amylin, and autopsy found that more than 90% of type Ⅱ diabetes patien...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/26C12N15/13C40B50/06
CPCC07K16/26C07K2317/56C40B50/06
Inventor 朱铁虹公倩李常颖畅继武甄云凤
Owner GENERAL HOSPITAL OF TIANJIN MEDICAL UNIV
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