CYP3A4 polypeptide

A peptide and amino acid technology, applied in the field of peptides

Active Publication Date: 2010-10-13
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 CYP3A4 polypeptide

Method used

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Experimental program
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specific Embodiment approach 1

[0009] Embodiment 1: The amino acid sequence of the CYP3A4 polypeptide in this embodiment is: SerGlnAsnSerLysGluThrGluSerHisLysAlaLeuSerAsp.

specific Embodiment approach 2

[0010] 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

[0011] Synthesis: synthesis and purification of peptide automatic synthesizer; (3) CYP3A4 polypeptide and carrier protein cross-linking: CYP3A4 peptide and carrier protein KLH are linked by glutaraldehyde, and then dialyzed with boric acid buffer solution with a pH value of 8.5 for 8 to 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 boost immunization repeatedly until the antibody titer is equal to or higher than 1:10000; (5) The serum containing the rabbit anti-human CYP3A4 polypepti...

specific Embodiment approach 3

[0013] 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 present invention relates to a polypeptide. 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 recombinantly expressed all amino acid antibodies for w...

Claims

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Application Information

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Patent Type & Authority Patents(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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