[Development of an anti-rTNF-alpha chimeric antibody].
OBJECTIVE
To develop an human-mouse chimeric antibody against rTNF-alpha.


METHODS
Expression vector of human-mouse chimeric antibody CZ12 gene was constructed by using VL and VH genes of mAb Z12 with neutralizing activity, and then COS7 cells were transfected. Supernatant containing CZ12 was detected by RT-PCR, ELISA, Western blot and in-vitro neutralization assay, respectively.


RESULTS
The RT-PCR analysis showed that there was the transcription of human-mouse chimeric antibody mRNA in the transfected COS7 cells. ELISA and Western blot detection showed that the CZ12 could recognize specifically TNF-alpha. The in-vitro neutralization assay proved that CZ12 could neutralize the cytoxicity of TNF-alpha to L929 cells.


CONCLUSIONS
human-mouse chimeric antibody CZ12 with neutralizing activity was successfully expressed in eukaryotic cells.
Lin
Zhou
Z
Basic Medical Institute,Academy of Military Medical Sciences, Beijing 100850,China.
Li
Yan
Y
Hu
Mei-Ru
MR
Yu
Ji-Yun
JY
Shen
Bei-Fen
BF
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D011993:Recombinant Fusion Proteins / Q000235:genetics; D014162:Transfection; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics
D000911:Antibodies, Monoclonal; D011993:Recombinant Fusion Proteins; D014409:Tumor Necrosis Factor-alpha