IgG light chain-independent secretion of heavy chain dimers: consequence for therapeutic antibody production and design

Stoyle, C. L., Stephens, P. E., Humphreys, D. P., Heywood, S., Cain, K. and Bulleid, N. J. (2017) IgG light chain-independent secretion of heavy chain dimers: consequence for therapeutic antibody production and design. Biochemical Journal, 474(18), pp. 3179-3188. (doi:10.1042/BCJ20170342) (PMID:28784690) (PMCID:PMC5590090)

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Abstract

Rodent monoclonal antibodies with specificity towards important biological targets are developed for therapeutic use by a process of humanisation. This process involves the creation of molecules, which retain the specificity of the rodent antibody but contain predominantly human coding sequence. Here we show that some humanised heavy chains can fold, form dimers and be secreted even in the absence of light chain. Quality control of recombinant antibody assembly in vivo is thought to rely upon folding of the heavy chain CH1 domain. This domain acts as a switch for secretion, only folding upon interaction with the light chain CL domain. We show that the secreted heavy-chain dimers contain folded CH1 domains and contribute to the heterogeneity of antibody species secreted during the expression of therapeutic antibodies. This subversion of the normal quality control process is dependent upon the heavy chain variable domain, is prevalent with engineered antibodies and can occur when only the Fab fragments are expressed. This discovery will impact on the efficient production of both humanised antibodies as well as the design of novel antibody formats.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Bulleid, Professor Neil and Stoyle, Dr Chloe
Authors: Stoyle, C. L., Stephens, P. E., Humphreys, D. P., Heywood, S., Cain, K., and Bulleid, N. J.
College/School:College of Medical Veterinary and Life Sciences > Institute of Molecular Cell and Systems Biology
Journal Name:Biochemical Journal
Publisher:Portland Press
ISSN:0264-6021
ISSN (Online):1470-8728
Published Online:07 August 2017
Copyright Holders:Copyright © 2017 The Authors
First Published:First published in Biochemical Journal 474(18):3179-3188
Publisher Policy:Reproduced under a Creative Commons License

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
617811Engineering the CHO host cell to improve protein productionNeil BulleidBiotechnology and Biological Sciences Research Council (BBSRC)BB/K501864/1RI MOLECULAR CELL & SYSTEMS BIOLOGY