[Construction of eukaryotic vector and expression of chimeric antibody against rh-bFGF].
OBJECTIVE
To express a human-mouse chimeric antibody against rh-bFGF antigen in eukaryotic cells.


METHODS
The VL and VH genes were amplified and cloned from the hybridoma 1F11 secreting anti-rh-bFGF mouse monoclonal antibody (mAb) and the CL and CH genes were amplified and cloned from the plasmid pMDHC and pMDLC. They were inserted into eukaryotic expression vectors after sequencing. The recombinant plasmids were then transformed into CHO-dhfr- cells for expression. The humanization and antigen specificity of expressed products were identified by indirect ELISA. The relative molecular mass (Mr) of expressed products was determined by SDS-PAGE.


RESULTS
The cloned antibody genes were identified to be functional by sequencing. The chimeric antibody could be detected in the culture supernatant of transformed CHO cells. The humanization and specificity of the chimeric antibody to rh-bFGF were confirmed by ELISA.


CONCLUSIONS
The mouse-human chimeric antibody against rh-bFGF has been expressed successfully in eukaryotic cells, which lays the foundation for its further clinical research.
Xiang
Jun-jian
JJ
Lab. of Molecular Immunology and Antibody Engineering, College of Life Sciences and Technology, Jinan University, Guangzhou 510632, China. txjj@jnu.edu.cn
Li
Hong-qing
HQ
Deng
Ning
N
Yang
Hong-yu
HY
Tong
Yi-gang
YG
Li
Jian-min
JM
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000235:genetics; D001483:Base Sequence; D016466:CHO Cells / Q000378:metabolism; D003001:Cloning, Molecular; D006224:Cricetinae; D016222:Fibroblast Growth Factor 2 / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D050438:Genes, Immunoglobulin Heavy Chain; D050439:Genes, Immunoglobulin Light Chain; D005822:Genetic Vectors; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics; D014170:Transformation, Genetic
D000911:Antibodies, Monoclonal; D011993:Recombinant Fusion Proteins; D016222:Fibroblast Growth Factor 2