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Updated: Aug 20, 2026
Laboratory Scale Production and Purification of a Therapeutic Antibody
Published on: January 24, 2017
[Construction and expression of anti-human CD3 chimeric antibody gene and preliminary study of expressed product]
Ming Lu1, Chun-Ning Lai, Ming Yu
1Institute of Basic Medical Sciences, Academy of Military Medical Sciences, Beijing 100850, China.
Aim:
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.
Conclusion:
The anti-human CD3 chimeric antibody has been constructed and expressed successfully, which lays the foundation for its further study.
