THE EFFECTS OF RECOMBINANT IGG4 EXPRESSION ON THE GROWTH OF SP2.0 CELL LINES

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JAHANDAR HODA; KHALAJ VAHID; NEMATOLLAHI LEILA; ZANDI FATEMEH; ENAYATI SOMAYEH; VAZIRI BEHROUZ

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Journal: Pathobiology Research Year:2014 | Volume:16 | Issue:4 Page(s): 27-37

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Title

THE EFFECTS OF RECOMBINANT IGG4 EXPRESSION ON THE GROWTH OF SP2.0 CELL LINES

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Keywords

MONOCLONAL ANTIBODYQ2

IGG4Q2

SP2.0 CELL LINEQ2

MAXIMUM CELL DENSITYQ2

Abstract

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-CLIghk construct.

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APA: Copy

JAHANDAR, HODA, KHALAJ, VAHID, NEMATOLLAHI, LEILA, ZANDI, FATEMEH, ENAYATI, SOMAYEH, & VAZIRI, BEHROUZ. (2014). THE EFFECTS OF RECOMBINANT IGG₄ EXPRESSION ON THE GROWTH OF SP2.0 CELL LINES. PATHOBIOLOGY RESEARCH (MODARES JOURNAL OF MEDICAL SCIENCES), 16(4), 27-37. SID. https://sid.ir/paper/81276/en

Vancouver: Copy

JAHANDAR HODA, KHALAJ VAHID, NEMATOLLAHI LEILA, ZANDI FATEMEH, ENAYATI SOMAYEH, VAZIRI BEHROUZ. THE EFFECTS OF RECOMBINANT IGG₄ EXPRESSION ON THE GROWTH OF SP2.0 CELL LINES. PATHOBIOLOGY RESEARCH (MODARES JOURNAL OF MEDICAL SCIENCES)[Internet]. 2014;16(4):27-37. Available from: https://sid.ir/paper/81276/en

IEEE: Copy

HODA JAHANDAR, VAHID KHALAJ, LEILA NEMATOLLAHI, FATEMEH ZANDI, SOMAYEH ENAYATI, and BEHROUZ VAZIRI, “THE EFFECTS OF RECOMBINANT IGG₄ EXPRESSION ON THE GROWTH OF SP2.0 CELL LINES,” PATHOBIOLOGY RESEARCH (MODARES JOURNAL OF MEDICAL SCIENCES), vol. 16, no. 4, pp. 27–37, 2014, [Online]. Available: https://sid.ir/paper/81276/en

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