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CYP3A4 polypeptide and preparation method of polypeptide antibody to human CYP3A4

A CYP3A4-KLH, polypeptide antibody technology, applied in the direction of anti-animal/human immunoglobulin, peptide source, animal/human peptide, etc., can solve the problems of high antibody cost, high CYP3A4 price, low safety and so on

Active Publication Date: 2007-02-28
HARBIN MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The purpose of the present invention is to solve the problem that the currently used human CYP3A4 is expensive, the cost of the prepared antibody is too high, and the safety is low, and a method for preparing a CYP3A4 polypeptide and an anti-human CYP3A4 polypeptide antibody is provided

Method used

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  • CYP3A4 polypeptide and preparation method of polypeptide antibody to human CYP3A4
  • CYP3A4 polypeptide and preparation method of polypeptide antibody to human CYP3A4
  • CYP3A4 polypeptide and preparation method of polypeptide antibody to human CYP3A4

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specific Embodiment approach 1

[0005] Embodiment 1: The amino acid sequence of the CYP3A4 polypeptide in this embodiment is: SerGlnAsnSerLysGluThrGluSerHisLysAlaLeuSerAsp.

specific Embodiment approach 2

[0006] Specific embodiment two: In this embodiment, the anti-human CYP3A4 polypeptide antibody is prepared according to the following steps: (1) Analysis of CYP3A4 epitope: use DNAStar software analysis to determine 15 amino acids at positions 278-292 of CYP3A4 protein as the amino acid sequence of the synthetic polypeptide; ( 2) Synthesis of CYP3A4 polypeptide: Synthesized and purified by an automatic polypeptide synthesizer; (3) Cross-linking of CYP3A4 polypeptide and carrier protein: CYP3A4 polypeptide and carrier protein KLH are linked by glutaraldehyde, and then boric acid buffer with a pH value of 8.5 Dialysis for 8-12 hours, cross-linking to form CYP3A4-KLH; (4) Preparation of rabbit anti-CYP3A4 polypeptide antibody: subcutaneously inject the emulsified CYP3A4-KLH on the back of the rabbit, and repeatedly boost the immunization until the antibody titer is equal to or higher than 1: 10000; (5) collecting and separating serum containing rabbit anti-human CYP3A4 polypeptide...

specific Embodiment approach 3

[0008] Specific embodiment three: the difference between this embodiment and specific embodiment two is: in step (two), a conventional C18 RP-HPLC column is used for purification. Other steps are the same as the second embodiment.

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Abstract

The invention discloses a preparing method to produce CYP3A4 polypeptide with amino acid sequence: SQNSKETESHKALSD and antihuman CYP3A4 polypeptide antibody, which comprises the following steps: (1)analysis of CYP3A4 antigen epitope; (2) synthesis of CYP3A4 polypeptide; (3)crosslinking of CYP3A4 polypeptide and carrier protein; (4) preparing antirabbit CYP3A4 polypeptide antibody; (5) assembling, separating to get serum with antibody; purifying antibody; getting antihuman CYP3A4 polypeptide antibody.

Description

technical field [0001] The invention relates to a polypeptide and a preparation method of the polypeptide antibody. Background technique [0002] Human CYP3A4 (CYP4503A4) is a member of the CYP3A subfamily, mainly expressed in the liver and small intestine. CYP3A4 is an important enzyme among human drug-metabolizing enzymes. Clinically, CYP3A4 can fully or partially metabolize more than half of the drugs. In particular, CYP3A4 is closely related to the metabolic transformation of exogenous carcinogens, anticancer drugs, and endogenous steroid hormones, vitamins, and fatty acids. Currently used human CYP3A4 (ab22702: http: / / www.abcam.com / provides goat anti-rabbit CYP3A4 synthetic peptide antibody for immunohistochemistry and western blot tests. Ab22704: http: / / www.abcam.com / Provide rabbit anti-human CYP3A4 artificially synthesized peptide antibody for immunohistochemistry and western blotting.Ab3572: http: / / www.abcam.com / provides rabbit anti-human CYP3A4 gene recombinant...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/435C07K16/18
Inventor 唐晓波朱大岭刘海英杨微武正华郭守利
Owner HARBIN MEDICAL UNIVERSITY
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