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Updated: Aug 15, 2026
Laboratory Scale Production and Purification of a Therapeutic Antibody
Published on: January 24, 2017
[Construction of anti-human VEGF165 chimeric antibodies and expression in eukaryotic cells]
1Cancer Institute (Hospital), Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing 100021.
Objective:
To express chimeric anti-VEGF165 antibody genes in eukaryotic cells.
Methods:
The variable region genes of light and heavy chains from mouse anti-human VEGF165 monoclonal antibody VmD11 were cloned into eukaryotic expression vectors and transfected into dihydrofolate reductase-deficient Chinese hamster ovary(CHO-dhfr-) cells to express chimeric antibody. The antibody expressed was examined for the presence of human constant regions and specificity against human VEGF165.
Results:
The chimeric antibody with human constant regions and specificity against human VEGF was detected in the culture supernatant of transfected CHO cells by ELISA and Western blot. The chimeric antibody gene was also detected at mRNA level by RT-PCR.
Conclusion:
The mouse anti-human VEGF165 monoclonal antibody is successfully humanized and expressed in eukaryotic cells.
