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Updated: Jul 14, 2026
Murine Model of CD40-activation of B cells
Published on: March 5, 2010
[Expression of human-mouse chimeric antibody against CD40 in CHO cell line and characterization of its function]
Qiu-xia Qu1, Cheng Chen, Qin Wang
1Key Laboratory of Clinical Immunology of Jiangsu Province, First Affiliated Hospital, Soochow University, Suzhou 215006, China.
Aim:
To investigate the stable expression of a chimeric antibody against CD40 moleculeèch-5C11éin CHO and its biological activity.
Methods:
Human-mouse chimeric antibody against CD40 recombinant plasmid and mock plasmid were transfected into CHO cell line through lipofectamine mediation. Human kappa chain and Fc fragment of ch-5C11 were characterized by FACS and Western blot. The concentration of ch-5C11 in cell supernatants was detected by Lowry assay and the inhibitory effect of ch-5C11 on the proliferation of Daudi cells was detected by MTT assay.
Results:
RT-PCR showed that target CHO cells integrated chimeric heavy chain and chimeric light chain gene. FACS and Western blot showed that ch-5C11 in cell supernatants maintained the binding activity and specificity to human CD40 molecule, and contained human kappa chain and Fc fragment. Cell supernatants were purified using protein G affinity chromatography. The concentration of human-mouse chimeric antibody against CD40 in cell supernatants was 0.535 mg/L. When co-cultured with B lymphoma cell line Daudi, ch-5C11 induced proliferation arrest of Daudi cells.
Conclusion:
The human-mouse chimeric antibody against CD40 can be expressed in CHO stably and effectively, which inhibits proliferation of Daudi.
