The Effects of Recombinant IgG۴ Expression on the Growth of Sp۲.۰ Cell Lines

Objective: The study of physiological changes in recombinant cell lines provides useful information to improve production performance. In this study, we investigate the effects of an anti-CD۳۳ chimeric IgG۴ expression on Sp۲.۰ cell growth. Methods: Variable region genes of light and heavy chains of monoclonal antibody produced by M۱۹۵ were cloned in pFUSE-CLIg-hk and pFUSE-CHIg-hG۴ expression vectors, respectively. Transfection of recombinant plasmids into Sp۲.۰ cell lines was performed using lipofectamine in two steps. Positive transformant cells were isolated and subjected to PCR, RT-PCR and Western blot analysis to confirm the integration of gene cassettes and the expression of recombinant IgG۴. To assess the growth parameters, recombinant and parent Sp۲.۰ cell lines were seeded at a density of ۱×۱۰۵ cells/ml in duplicate into ۱۲-well plates. For nine days, culture plates were sampled daily and viable cell count and viability determined. Results: The results of PCR, RT-PCR and Western blot analyses confirmed the generation of stable producer cell lines. In recombinant cells, the maximum cell density decreased by ۴۶%. However, it was observed that IgG۴ expression had no effect on cell viability of these transfectants. Conclusion: Our results showed that the expression of recombinant IgG۴ can change growth parameters in Sp۲.۰ cell lines that express the pFUSE-CHIg-hG۴-pFUSE-CLIg-hk construct.

کلیدواژه ها:

IgG۴ ، Monoclonal antibody ، IgG۴ ، Sp۲.۰ cell line ، Maximum cell density ، آنتیبادی تک دودمانی ، سلول Sp۲.۰ ، حداکثر چگالی سلولی