Stable expression of targeting complement inhibitor CR2-CD59 in Chinese hamster ovary cells
- VernacularTitle:人源靶向补体抑制物CR2-CD59在中国仓鼠卵巢细胞中的稳定表达
- Author:
Yan GUO
;
Zhihua KOU
;
Shihui SUN
;
Chuanfu ZHANG
;
Guangyu ZHAO
;
Hong YU
;
Hongbin SONG
;
Fei QIAO
;
Wanrong CHEN
;
Yusen ZHOU
- Publication Type:Journal Article
- Keywords:
Chinese hamster ovary cell;
CR2-CD59;
complement
- From:Journal of Third Military Medical University
2003;0(09):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To obtain Chinese hamster ovary (CHO) cell lines that stably express a targeting complement inhibitor CR2-CD59.Methods The recombinant plasmid PEE14.1-CR2-CD59 was constru-cted by cloning the DNA fragment CR2-CD59 into plasmid PEE14.1,and the obtained plasmid was transfected into CHO cells by FuGENE 6.The clones with stable high expression of target fragment were selected by methionine sulfoximine (MSX),the expression of CR2-CD59 was analyzed by ELISA,SDS-PAGE and Western blotting analysis.Results Several stable expression clones were obtained,and CR2-CD59 was highly expressed in the secret form in CHO cells.SDS-PAGE analysis showed that the molecular weight of the recombined protein CR2-CD59 was consistent with the predicted one.ELISA and Western blotting results revealed that the CR2-CD59 could react with both anti-human CR2 and anti-human CD59 polyclonal antibodies.Compared with serum-containing medium,the protein was highly expressed in serum-free medium (P
