[Construction of GPI-CTLA4Ig chimeric molecule and its expression on CHO-dhfr(-) cells].
OBJECTIVE
To develop a novel immunosuppressant GPI-CTLAIg modified by glycosyl-phosphatidylinositol(GPI).


METHODS
GPI-modified CTLAIg was produced by linking-up of CTLA4Ig with GPI-modification signal sequences from decay-accelerating factor (DAF). Chimeric molecule GPI-CTLA4Ig gene was cloned into eukaryotic expression vector pCI-dhfr. Using lipofectine-mediated gene transfer technique, pCI-GPI-CTLA4Ig was transfected into CHO-dhfr(-) cells, and the transfectants were screened by methotrexate (MTX). Expression of the recombinant protein was assessed by RT-PCR, ELISA, cell immunofluorescence staining and Western blot, and the purification of expressed protein was performed by protein A affinity chromatography.


RESULTS
The chimeric molecule GPI-CTLA4Ig has been constructed and stablely expressed on CHO-dhfr(-) cells.


CONCLUSIONS
GPI-modified CTLAIg will may be used as novel immunosuppressant for suppressing reaction in graft rejection.
Yu
Ji-Yun
JY
Institute of Basic Medical Sciences, Academy of Military Medical Sciences, Beijing 100850,China.
Shen
Bei-Fen
BF
Li
Yan
Y
Chen
Xing
X
Cheng
Shao-Hui
SH
Tian
Rong
R
Qu
Mei-Hua
MH
D000818:Animals; D018958:CD55 Antigens; D016466:CHO Cells; D018014:Gene Transfer Techniques; D006801:Humans; D011993:Recombinant Fusion Proteins / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics; D014162:Transfection
D018958:CD55 Antigens; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins