Expression of ATR-Fc fusion protein in CHO cells.
- Author:
Li-Hua GAO
1
;
Xian-Wen HU
;
Wei CHEN
;
Jun-Jie XU
;
Jian ZHAO
;
Hui-Peng CHEN
Author Information
1. Beijing Institute of Biotechnology, Beijing 100071, China.
- Publication Type:Journal Article
- MeSH:
Animals;
CHO Cells;
Cricetinae;
Cricetulus;
Gene Transfer Techniques;
Genetic Vectors;
Humans;
Immunoglobulin Fc Fragments;
biosynthesis;
genetics;
Immunoglobulin G;
biosynthesis;
genetics;
Neoplasm Proteins;
biosynthesis;
genetics;
Receptors, Cell Surface;
biosynthesis;
genetics;
Recombinant Fusion Proteins;
biosynthesis;
genetics;
immunology
- From:
Chinese Journal of Biotechnology
2005;21(5):826-831
- CountryChina
- Language:Chinese
-
Abstract:
ATR-Fc is a fusion protein consisting of extracellular domain of human anthrax toxin receptor (ATR) and a fragment (hinge, CH2, and CH3 domains) of the Fc of human IgG1. The aim of ATR-Fc expression is to get an antibody-like molecule binding to protective antigen (PA), a component of anthrax toxins, this fusion protein may compete with cell surface receptor for PA binding, and block the transport of lethal factor (LF) and edema factor (EF) into cells, thereby act as an antitoxin to prevent and treat anthrax infection. A DNA fragment encoding N-terminal amino acids 1-227 of ATR and human IgG1 Fc was inserted into the Hind III and Not I sites of pcDNA3.1 to generate the eukaryotic vector pcDNA3.1/ATR-Fc for expression of ATR-Fc fusion protein. Using lipofectine-mediated gene transfer technique, pcDNA3.1/ATR-Fc was transfected into CHO-K1 cells. After selected with G418, a recombinant CHO cell line, ATR-Fc-1D5, whose expression level was about 10 - 15 microg/(10(6) cells x d), was established. The recombinant protein expressed by the ATR-Fc-1D5 cells was purified with protein A chromatography. The experimental results demonstrated a direct and specific interaction between ATR-Fc and PA assessed by ELISA.
