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2
Optimization of heavy chain and light chain signal peptides for high level expression of therapeutic antibodies in CHO cells.优化重链和轻链信号肽以实现治疗性抗体在CHO细胞中的高水平表达。
PLoS One. 2015 Feb 23;10(2):e0116878. doi: 10.1371/journal.pone.0116878. eCollection 2015.
3
Optimization of chemically defined feed media for monoclonal antibody production in Chinese hamster ovary cells.优化用于中国仓鼠卵巢细胞中单克隆抗体生产的化学成分确定的培养基。
J Biosci Bioeng. 2015 Jul;120(1):78-84. doi: 10.1016/j.jbiosc.2014.11.022. Epub 2015 Feb 10.
4
Rational design of viscosity reducing mutants of a monoclonal antibody: hydrophobic versus electrostatic inter-molecular interactions.单克隆抗体减粘突变体的合理设计：疏水与静电分子间相互作用
MAbs. 2015;7(1):212-30. doi: 10.4161/19420862.2014.985504.
5
Optimal charged mutations in the complementarity-determining regions that prevent domain antibody aggregation are dependent on the antibody scaffold.在互补决定区中进行最优的带电突变以防止结构域抗体聚集，这取决于抗体支架。
Protein Eng Des Sel. 2014 Feb;27(2):29-39. doi: 10.1093/protein/gzt058. Epub 2014 Jan 6.
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Proteins improving recombinant antibody production in mammalian cells.改善哺乳动物细胞中重组抗体生产的蛋白质。
Appl Microbiol Biotechnol. 2014 Feb;98(3):1031-42. doi: 10.1007/s00253-013-5427-3. Epub 2013 Dec 11.
7
Optimized signal peptides for the development of high expressing CHO cell lines.优化信号肽以开发高表达 CHO 细胞系。
Biotechnol Bioeng. 2013 Apr;110(4):1164-73. doi: 10.1002/bit.24776. Epub 2013 Jan 17.
8
Progress in overcoming the chain association issue in bispecific heterodimeric IgG antibodies.克服双特异性异二聚体 IgG 抗体中链关联问题的进展。
MAbs. 2012 Nov-Dec;4(6):653-63. doi: 10.4161/mabs.21379. Epub 2012 Aug 27.
9
Production of antibody fragments using the baculovirus-insect cell system.利用杆状病毒-昆虫细胞系统生产抗体片段。
Methods Mol Biol. 2012;907:371-87. doi: 10.1007/978-1-61779-974-7_22.
10
Production of antibody fragments in Escherichia coli.在大肠杆菌中生产抗体片段。
Methods Mol Biol. 2012;907:305-24. doi: 10.1007/978-1-61779-974-7_18.

重链和轻链之间界面的静电工程促进抗体Fab片段的产生。

Electrostatic engineering of the interface between heavy and light chains promotes antibody Fab fragment production.

作者信息

Ohmuro-Matsuyama Yuki, Mori Keita, Hamada Hirotsugu, Ueda Hiroshi, Yamaji Hideki

机构信息

Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, 1-1 Rokkodai, Nada, Kobe, 657-8501, Japan.

Manufacturing Technology Association of Biologics (MAB), c/o Center for Business Incubation, Kobe University, 1-5-6 Minatojima-Minamimachi, Chuo-ku, Kobe, 650-0047, Japan.

出版信息

Cytotechnology. 2017 Jun;69(3):469-475. doi: 10.1007/s10616-016-9955-4. Epub 2016 Feb 8.

DOI:10.1007/s10616-016-9955-4
PMID:26856589
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5461237/
Abstract

Monoclonal antibodies and antibody fragments are used for diverse diagnostic and therapeutic applications. We have investigated the secretory production of Fab fragments from insect cells cotransfected with plasmid vectors carrying heavy- and light-chain genes. In the present study, to promote the formation of the disulfide bond between the heavy and light chains, some positively charged amino acid residues were introduced near the cysteine residue for the disulfide bond at the C-terminus of C, while some negatively charged amino acid residues were added near the cysteine residue for the disulfide bond at the C-terminus of C. This electrostatic steering led to an increase in Fab secretions from insect cells.

摘要

单克隆抗体和抗体片段被用于多种诊断和治疗应用。我们研究了用携带重链和轻链基因的质粒载体共转染的昆虫细胞中Fab片段的分泌产生。在本研究中，为促进重链和轻链之间二硫键的形成，在C末端形成二硫键的半胱氨酸残基附近引入了一些带正电荷的氨基酸残基，同时在C末端形成二硫键的半胱氨酸残基附近添加了一些带负电荷的氨基酸残基。这种静电引导导致昆虫细胞中Fab分泌增加。