[Construction and expression of anti-human CD3 chimeric antibody gene and preliminary study of expressed product].
OBJECTIVE
To construct and express anti-human CD3 chimeric antibody.


METHODS
The genes of variable regions of the light chain (V(L)) and heavy chain (V(H)) were cloned respectively into the expression vectors (V(L) Express, V(H) Express), and co-transfected into COS-7 cells. Expression level of the chimeric antibody in culture supernatant was detected by ELISA. The antibody was purified through protein A affinity chromatography and identified by Western blot. Binding activity of the chimeric antibody to the antigen was determined by FACS. Biological activity of the chimeric antibody was determined by mixed T-lymphocyte culture test.


RESULTS
The expression vectors were constructed and the anti-human CD3 chimeric antibody was expressed and purified successfully. Western blot showed that the purified antibody was human-mouse chimeric antibody. FACS result showed that the chimeric antibody had antigen-binding activity. Mixed T-lymphocyte culture test showed that the chimeric antibody could suppress proliferation of T lymphocytes.


CONCLUSIONS
The anti-human CD3 chimeric antibody has been constructed and expressed successfully, which lays the foundation for its further study.
Lu
Ming
M
Institute of Basic Medical Sciences, Academy of Military Medical Sciences, Beijing 100850, China.
Lai
Chun-Ning
CN
Yu
Ming
M
Guo
Ning
N
Li
Yan
Y
Shen
Bei-Fen
BF
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology* / Q000302:isolation & purification; D015153:Blotting, Western; D017252:CD3 Complex / Q000276:immunology*; D019556:COS Cells; D049109:Cell Proliferation; D002522:Chlorocebus aethiops; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D008214:Lymphocytes / Q000166:cytology / Q000276:immunology; D051379:Mice; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology* / Q000302:isolation & purification
D000911:Antibodies, Monoclonal; D017252:CD3 Complex; D011993:Recombinant Fusion Proteins