Preparation of anfi-CD3 and anti-IgM&mgr; chain bispecific antibody by cell fusion.
OBJECTIVE: To prepare anti-CD3 and anti-Igm&mgr; chain bispecific antibody (BsAb) by means of cell fusion and assess the stability and activity of the hybrid hybridoma. METHODS: Mouse hybridoma cell line secreting anti-human CD3 monoclonal antibody (mAb) was trans formed into HAT-sensitive cells by 8-azaguanine, which was subsequently transfected by plasmid pCDaA3 containing neoR marker gene via FuGENETM6. The resulted mutant phenotype, named alphaCD3 HATs G418R, was fused with the hybridoma producing anti-IgM&mgr; chain mAb, and enzyme-linked immunosorbent assay and flow cytometry were employed to identify the fusion cells producing the target BsAb. RESULTS: Six rounds of cell fusion was performed and a total of l 080 wells inoculated, yielding 5 hybrid hybridoma cell lines. Two of the 5 cell lines were subcloned and continuously cultured in vitro for 2 months without losing their capability to secrete BsAb. CONCLUSION: Cell fusion technique can be utilized to prepare BsAb-producing hybrid hybridoma that has similar stability and activity to its parent hybridoma.
Su
Jin
J
Department of Respiratory Diseases, Nanfang Hospital, First Military Medical University, Guangzhou 510515, China.
Zhang
Ya-Li
YL
Guo
Zhi-Bing
ZB
Che
Xiao-Yan
XY
Wang
Xiao-Ning
XN