Literature DB >> 26856589

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

Yuki Ohmuro-Matsuyama1,2, Keita Mori1, Hirotsugu Hamada1, Hiroshi Ueda3, Hideki Yamaji4,5.   

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 CL, while some negatively charged amino acid residues were added near the cysteine residue for the disulfide bond at the C-terminus of CH1. This electrostatic steering led to an increase in Fab secretions from insect cells.

Entities:  

Keywords:  Antibody; Charged amino acid; Electrostatic steering; Fab fragment; Insect cell

Year:  2016        PMID: 26856589      PMCID: PMC5461237          DOI: 10.1007/s10616-016-9955-4

Source DB:  PubMed          Journal:  Cytotechnology        ISSN: 0920-9069            Impact factor:   2.058


  31 in total

1.  Protein aggregation during overexpression limited by peptide extensions with large net negative charge.

Authors:  Yian-Biao Zhang; Jason Howitt; Sean McCorkle; Paul Lawrence; Karen Springer; Paul Freimuth
Journal:  Protein Expr Purif       Date:  2004-08       Impact factor: 1.650

2.  Optimal charged mutations in the complementarity-determining regions that prevent domain antibody aggregation are dependent on the antibody scaffold.

Authors:  Joseph M Perchiacca; Christine C Lee; Peter M Tessier
Journal:  Protein Eng Des Sel       Date:  2014-01-06       Impact factor: 1.650

3.  Rational design of viscosity reducing mutants of a monoclonal antibody: hydrophobic versus electrostatic inter-molecular interactions.

Authors:  Pilarin Nichols; Li Li; Sandeep Kumar; Patrick M Buck; Satish K Singh; Sumit Goswami; Bryan Balthazor; Tami R Conley; David Sek; Martin J Allen
Journal:  MAbs       Date:  2015       Impact factor: 5.857

Review 4.  Progress in overcoming the chain association issue in bispecific heterodimeric IgG antibodies.

Authors:  Christian Klein; Claudio Sustmann; Markus Thomas; Kay Stubenrauch; Rebecca Croasdale; Jürgen Schanzer; Ulrich Brinkmann; Hubert Kettenberger; Jörg T Regula; Wolfgang Schaefer
Journal:  MAbs       Date:  2012-08-27       Impact factor: 5.857

5.  Optimized signal peptides for the development of high expressing CHO cell lines.

Authors:  Lars Kober; Christoph Zehe; Juergen Bode
Journal:  Biotechnol Bioeng       Date:  2013-01-17       Impact factor: 4.530

6.  VH/VL interface engineering to promote selective expression and inhibit conformational isomerization of thrombopoietin receptor agonist single-chain diabody.

Authors:  Tomoyuki Igawa; Hiroyuki Tsunoda; Yasufumi Kikuchi; Maki Yoshida; Megumi Tanaka; Akiko Koga; Yasuo Sekimori; Tetsuro Orita; Yoshinori Aso; Kunihiro Hattori; Masayuki Tsuchiya
Journal:  Protein Eng Des Sel       Date:  2010-06-24       Impact factor: 1.650

7.  Enhanced periplasmic expression of high affinity humanized scFv against Hepatitis B surface antigen by codon optimization.

Authors:  Ashutosh Tiwari; Anurag Sankhyan; Navin Khanna; Subrata Sinha
Journal:  Protein Expr Purif       Date:  2010-06-20       Impact factor: 1.650

8.  A comparative study of different vector designs for the mammalian expression of recombinant IgG antibodies.

Authors:  Jiandong Li; Christian Menzel; Doris Meier; Congcong Zhang; Stefan Dübel; Thomas Jostock
Journal:  J Immunol Methods       Date:  2006-11-13       Impact factor: 2.303

Review 9.  Metabolic flux analysis in mammalian cell culture.

Authors:  Lake-Ee Quek; Stefanie Dietmair; Jens O Krömer; Lars K Nielsen
Journal:  Metab Eng       Date:  2009-10-13       Impact factor: 9.783

10.  Optimization of heavy chain and light chain signal peptides for high level expression of therapeutic antibodies in CHO cells.

Authors:  Ryan Haryadi; Steven Ho; Yee Jiun Kok; Helen X Pu; Lu Zheng; Natasha A Pereira; Bin Li; Xuezhi Bi; Lin-Tang Goh; Yuansheng Yang; Zhiwei Song
Journal:  PLoS One       Date:  2015-02-23       Impact factor: 3.240

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  1 in total

1.  Modifications of a signal sequence for antibody secretion from insect cells.

Authors:  Yuki Ohmuro-Matsuyama; Hideki Yamaji
Journal:  Cytotechnology       Date:  2017-06-05       Impact factor: 2.058

  1 in total

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