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