Method for separating membrane receptor protein

A receptor protein and separation membrane technology, applied in peptide preparation methods, chemical instruments and methods, animal/human proteins, etc., can solve problems such as cumbersome operations, single binding protein bands, and long time required

Inactive Publication Date: 2015-04-08
敖云霞
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In the past 10 years, scholars at home and abroad have used GST pull-down technology, co-immunoprecipitation and phage display technology to screen the protein bound to preS1 by liver cell membrane protein. The binding protein band is relatively single

Method used

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Embodiment Construction

[0008] The present invention will be described in further detail below, but not as any limitation to the present invention.

[0009] Embodiments of the invention:

[0010] 1 Materials and methods

[0011] 1.1 Materials and instruments

[0012] HepG2 human liver cancer cells were purchased from the Cell Bank of the Chinese Academy of Sciences; the membrane protein extraction kit was purchased from Beijing Pulilai Biotechnology Company; ; streptavidin-labeled magnetic beads were purchased from Baihuishen Biotechnology Co., Ltd.; DMEM high-glucose medium and trypsin were purchased from Hyclone Company of the United States; Company), DYY-Ⅲ-5 steady flow electrophoresis apparatus and electrophoresis tank (Beijing Liuyi Instrument Factory), UMAX scanner (UMAX company).

[0013] 1.2 Method

[0014] 1.2.1 Isolation of preS1-binding protein by immunomagnetic beads

[0015] Basic operation steps: (1) Cell culture collection, HepG2 cells were cultured in DMEM high-glucose medium con...

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Abstract

The invention discloses a method for separating a membrane receptor protein. The protein combined with a preS1 peptide fragment of a hepatitis B virus on a cytomembrane of a hepatoma carcinoma cell is separated by virtue of immunomagnetic beads; and a method for rapidly separating the membrane receptor protein is built. According to the research result, the molecular weight range of a band is roughly equivalent to that of the foregoing scholar; the band is relatively abundant; the immunomagnetic beads are feasible for separating receptor associated protein; the overall process from preparation of the membrane protein to gel scanning can be finished within 2d; and the operation is very simple. Compared with other methods, according to the method disclosed by the invention, the effects of saving time and labor can be achieved, and the method is convenient and fast.

Description

technical field [0001] The invention relates to a method for separating membrane receptor proteins, belonging to the technical field of biopharmaceuticals. Background technique [0002] Hepatitis B virus (HBV) is a pathogen that causes chronic hepatitis B in humans, and its mechanism of infecting liver cells is still unclear. It is generally believed that HBV may interact with the corresponding receptor protein on the liver cell membrane through its envelope protein. Most of the known HBV candidate receptors interact with the 21-47 amino acids in the preS1 region; Abou-Jaoud et al. proved the importance of the preS1 region in HBV infection. In the past 10 years, scholars at home and abroad have used GST pull-down technology, co-immunoprecipitation and phage display technology to screen the protein bound to preS1 by liver cell membrane protein. The binding protein band is relatively single. Contents of the invention [0003] The purpose of the present invention is to prov...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/02C07K1/26
CPCC07K14/705Y02P20/10Y02P20/50
Inventor 敖云霞
Owner 敖云霞
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