[Construction of expression vector for human-mouse chimeric antibody and expression of an antibody against human HER2].
OBJECTIVE
To construct universal eukaryotic expression vector for expressing human-mouse chimeric antibody and have application in expression of PCR-obtained mouse variable region gene fragments in the form of human-mouse chimeric antibody, for the purpose of clinical therapy.


METHODS
By using human Tac antigen signal peptide and kappa chain and gamma 1 heavy chain gene fragments of human Ig, we constructed a versatile expression vector for human-mouse chimeric antibody and used it to transfect cultured 293T cells. The expressed product was detected using RT-PCR, FACS and ELISA.


RESULTS
An expression vector for PCR-derived V genes of mouse Ig as human-mouse chimeric antibody was constructed. The V(L) and V(H) genes encoding mouse anti-human HER2 antibody were amplified with primers designed for this expression system, and an expression vector for chimeric antibody against human HER2 was constructed. By transfection of 293T cells, we demonstrated that a chimeric antibody was produced and secreted to the culture medium.


CONCLUSIONS
This expression system is useful for antibody engineering for it provides a simple way to express chimeric antibody from V genes of mouse antibody obtained by PCR.
Wang
Li-li
LL
Institute of Modern Separation Sciences, Northwest University, Xi'an 710069, China.
Han
Hua
H
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D002460:Cell Line; D004622:Embryo, Mammalian; D004847:Epithelial Cells / Q000378:metabolism; D018734:Genes, erbB-2; D005822:Genetic Vectors; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D007147:Immunoglobulin Light Chains / Q000235:genetics; D007135:Immunoglobulin Variable Region / Q000235:genetics*; D007142:Immunoglobulin gamma-Chains / Q000235:genetics; D007668:Kidney / Q000166:cytology; D051379:Mice; D021382:Protein Sorting Signals / Q000235:genetics; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000276:immunology*; D015375:Receptors, Interleukin-2 / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000276:immunology; D014162:Transfection
D000911:Antibodies, Monoclonal; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007135:Immunoglobulin Variable Region; D007142:Immunoglobulin gamma-Chains; D021382:Protein Sorting Signals; D018719:Erb-b2 Receptor Tyrosine Kinases; D015375:Receptors, Interleukin-2; D011993:Recombinant Fusion Proteins