[Construction of anti-human VEGF165 chimeric antibodies and expression in eukaryotic cells].
OBJECTIVE
To express chimeric anti-VEGF165 antibody genes in eukaryotic cells.


METHODS
The variable region genes of light and heavy chains from mouse anti-human VEGF165 monoclonal antibody VmD11 were cloned into eukaryotic expression vectors and transfected into dihydrofolate reductase-deficient Chinese hamster ovary(CHO-dhfr-) cells to express chimeric antibody. The antibody expressed was examined for the presence of human constant regions and specificity against human VEGF165.


RESULTS
The chimeric antibody with human constant regions and specificity against human VEGF was detected in the culture supernatant of transfected CHO cells by ELISA and Western blot. The chimeric antibody gene was also detected at mRNA level by RT-PCR.


CONCLUSIONS
The mouse anti-human VEGF165 monoclonal antibody is successfully humanized and expressed in eukaryotic cells.
Ran
Y
Y
Cancer Institute (Hospital), Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing 100021.
Yang
Z
Z
Wang
G
G
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D015153:Blotting, Western; D016466:CHO Cells; D006224:Cricetinae; D016228:Endothelial Growth Factors / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D008222:Lymphokines / Q000276:immunology*; D051379:Mice; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis*; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors
D000911:Antibodies, Monoclonal; D016228:Endothelial Growth Factors; D008222:Lymphokines; D011993:Recombinant Fusion Proteins; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors