[Preparation of monoclonal antibodies to human vascular endothelial growth factor(VEGF121) and identification of its expression on gastric carcinoma cell line MGC803].
OBJECTIVE
To further elucidate the source of VEGF in solid tumors.


METHODS
Traditional hybridoma technology was used to prepare VEGF monoclonal antibodies (McAb); it's activity was measured by 3H-TdR incorporation on human umbilical vein endothelial cells (HUVEC). VEGF gene was amplified by PCR with cDNA as template from MGC803 cell lines. The PCR products were cloned into fusion protein prokaryotic expression vector PGEX2T which expressed in E. coli XL-1 blue. Western blot analysis was used to identify expression of VEGF.


RESULTS
The McAbs could specifically bind to VEGF in ELISA. One of the McAbs(5C5) could neutralize the mitogenic activity of VEGF121 on HUVEC in a dose-dependent manner. The VEGF gene fragment obtained from RT-PCR cloned to PGEX2T vector expressed a protein product which reacted specifically with McAb 5C5.


CONCLUSIONS
Gastric carcinoma is shown to express VEGF. It may be a major source of VEGF in human solid tumor. Monoclonal antibody against VEGF may be of value in the treatment of cancer.
Tian
X
X
Department of Biochemistry and Molecular Biology, School of Oncology, Beijing Medical University, 100034.
Wu
J
J
Meng
L
L
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology*; D015153:Blotting, Western; D016228:Endothelial Growth Factors / Q000032:analysis / Q000096:biosynthesis / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D008222:Lymphokines / Q000032:analysis / Q000096:biosynthesis / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D013274:Stomach Neoplasms / Q000737:chemistry*; D014407:Tumor Cells, Cultured; 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