文献类型：期刊文献

英文题名：Heavy chain variants affect light and heavy chains assembly of monoclonal antibody expressed by Pichia pastoris

作者：Gong, Xiulong[1];Ye, Kaixiong[1];Xu, Mingqiang[1];Qian, Zhilan[1];Liu, Qi[1];Wang, Xiaolong[2];Zhou, Xiangshan[2];Liu, Haifeng[3];Cai, Menghao[1,4]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, 130 Meilong Rd, Shanghai 200237, Peoples R China;[2]China Resources Biopharmaceut Co Ltd, Shenzhen, Peoples R China;[3]China Resources Angde Biotech Pharma Co Ltd, Liaocheng, Peoples R China;[4]Shanghai Collaborat Innovat Ctr Biomfg, Shanghai, Peoples R China

年份：2025

卷号：55

期号：8

起止页码：1009

外文期刊名：PREPARATIVE BIOCHEMISTRY & BIOTECHNOLOGY

收录：;WOS:【SCI-EXPANDED(收录号:WOS:001432805200001)】；

基金：This study was supported by National Key Research and Development Program of China (2022YFC2805102), Young Scientist Fund of National Natural Science Foundation of China (32201206), and China Postdoctoral Science Foundation (2022M711146).

语种：英文

外文关键词：Assembly; monoclonal antibody; Pichia pastoris; recombinant expression

摘要：Monoclonal Antibody accounts for the largest share of recombinant protein drugs and is the primary choice for the treatment of various diseases. In this study, the monoclonal antibody Eptinezumab was expressed by a yeast host Pichia pastoris. Although the expression and secretion of light chain was efficient, the assembly efficiency between light and heavy chains was low. As the retention of the heavy chain in endoplasmic reticulum may trigger protein degradation, ERAD ubiquitination-related genes were then knocked out separately but it only led to minor improvement effect. Expression of splitted heavy chain variants further revealed that although endoplasmic reticulum retention of the heavy chain upregulated KAR2 expression, it did not affect the assembly efficiency of the light and heavy chains. It was inferred that binding of complete heavy chains to KAR2 spatially affected the assembly between light and heavy chains. Design and screening of KAR2 variants that facilitating full-length antibody assembly could be preferentially considered in future.