Volume 16, Issue 4 (2014)                   mjms 2014, 16(4): 27-37 | Back to browse issues page

XML Persian Abstract Print

Download citation:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:
Jahandar H, Khalaj V, Nematollahi L, Zandi F, Enayati S, Vaziri B. The Effects of Recombinant IgG4 Expression on the Growth of Sp2.0 Cell Lines. mjms 2014; 16 (4) :27-37
URL: http://mjms.modares.ac.ir/article-30-12218-en.html
The Effects of Recombinant IgG4 Expression on the Growth of Sp2.0 Cell Lines
Hoda Jahandar1 , Vahid Khalaj1 , Leila Nematollahi1 , Fatemeh Zandi1 , Somayeh Enayati1 , Behrouz Vaziri 1
1- Department of Medical Biotechnology, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran
Abstract:   (11044 Views)
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-CD33 chimeric IgG4 expression on Sp2.0 cell growth. Methods: Variable region genes of light and heavy chains of monoclonal antibody produced by M195 were cloned in pFUSE-CLIg-hk and pFUSE-CHIg-hG4 expression vectors, respectively. Transfection of recombinant plasmids into Sp2.0 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 IgG4. To assess the growth parameters, recombinant and parent Sp2.0 cell lines were seeded at a density of 1×105 cells/ml in duplicate into 12-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 46%. However, it was observed that IgG4 expression had no effect on cell viability of these transfectants. Conclusion: Our results showed that the expression of recombinant IgG4 can change growth parameters in Sp2.0 cell lines that express the pFUSE-CHIg-hG4-pFUSE-CLIg-hk construct.
Keywords: IgG4, Monoclonal antibody, IgG4, Sp2.0 cell line, Maximum cell density
Full-Text [PDF 943 kb]   (6363 Downloads)    
Article Type: Original Manuscipt | Subject: Biotechnology
Received: 2014/01/5 | Accepted: 2014/02/2
Rights and permissions
Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.