文献类型：期刊文献

英文题名：Limitations in the process of transcription and translation inhibit recombinant human chorionic gonadotropin expression in CHO cells

作者：Liu, Yang[1];Yi, Xiaoping[1];Zhuang, Yingping[1];Zhang, Siliang[1]

机构：[1]E China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China

年份：2015

卷号：204

起止页码：63

外文期刊名：JOURNAL OF BIOTECHNOLOGY

收录：;EI(收录号：20151900818619);WOS:【SCI-EXPANDED(收录号:WOS:000354400000013)】；

基金：This work was supported by grants from the Major State Basic Research Development Program of China (2013CB733600) and the Chinese National Programs for High Technology Research and Development (2012AA021201).

语种：英文

外文关键词：Transcription limitation; Recombinant protein; Mammalian cell culture

摘要：Human chorionic gonadotropin (hCG) is a glycoprotein hormone that exists as a heterodimer with a alpha subunit and beta subunit assembled together with disulfide bridges. This hormone plays an important role in the detection of ovulation induction and in the treatment of certain diseases that cause female infertility. The effects of transcription, subunit expression, assembling and secretion on recombinant hCG expression in CHO cells were studied using stable high-producing and low-producing cell lines generated by the FLPIflTM system. The results indicated that the mRNA and polypeptide levels of the beta subunit were always higher than those of the ce. subunit. Further study confirmed that the differences were caused by the transcription rate rather than by mRNA stability. In the high-producing cell lines, there was obvious transcription level limitation of the ce. subunit in contrast to the beta subunit. In addition, there was obvious limitation of the synthetic steps from mRNA to polypeptide for both the ce. subunit and the beta subunit, especially the beta subunit. Significant limitations of the assembly and secretion levels were not observed in this research. This study presents a research methodology for double subunit protein expression and provides valuable evidence for the enhancement of recombinant hCG productivity. (C) 2014 Elsevier B.V. All rights reserved.