Comparison of Human Factor IX Abundance and mRNA Expression Levels In Stable Insect and Mammalian Cells

J. Vatandoost, Shadi Tirdad
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引用次数: 0

Abstract

In the production of recombinant proteins, the selection of an expression system is very important. Although CHO cells are used as mammalian expression system, the ability of insect Schneider line 2 (S2) cells as a new expression system for the high production of many human proteins were confirmed. It is suggested that high copy number of an introduced gene and so high transcription in these cells can be regarded as one of the reasons for high expression of recombinant proteins. Therefore, the present study aimed to evaluate the correlation of recombinant human coagulation factor IX (hFIX) abundance and mRNA expression. The amount of hFIX mRNA by quantitative real-time PCR as well as hFIX protein in cell lysate and cultured media by Elisa was analyzed. The results of data analysis indicated 6 fold increases in mRNA level in S2 cells in comparison to CHO cells. Furthermore, S2 cell line indicated 5.5 and 7-fold increase in total and secreted protein level, respectively, compared to CHO cell line. The data demonstrated the correlation of mRNA and protein abundance and indicate that S2 cell lines are superior in producing the recombinant proteins.
人因子IX在稳定昆虫和哺乳动物细胞中丰度和mRNA表达水平的比较
在重组蛋白的生产中,表达系统的选择是非常重要的。尽管CHO细胞被用作哺乳动物的表达系统,但昆虫Schneider系2(S2)细胞作为一种新的表达系统用于大量生产人类蛋白质的能力已得到证实。这表明,导入基因的高拷贝数和在这些细胞中的高转录可以被认为是重组蛋白高表达的原因之一。因此,本研究旨在评估重组人凝血因子IX(hFIX)丰度与mRNA表达的相关性。通过实时定量PCR分析hFIX mRNA的量,并通过Elisa分析细胞裂解物和培养基中hFIX蛋白的量。数据分析的结果表明,与CHO细胞相比,S2细胞中的mRNA水平增加了6倍。此外,与CHO细胞系相比,S2细胞系的总蛋白水平和分泌蛋白水平分别增加了5.5倍和7倍。这些数据证明了mRNA和蛋白质丰度的相关性,并表明S2细胞系在产生重组蛋白质方面是优越的。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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