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Tuning Relative Polypeptide Expression to Optimize Assembly, Yield and Downstream Processing of Bispecific Antibodies
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Antibodies 2018, 7(3), 35; https://doi.org/10.3390/antib7030035

Relative Contribution of Framework and CDR Regions in Antibody Variable Domains to Multimerisation of Fv- and scFv-Containing Bispecific Antibodies

Protein Sciences Group, UCB Pharma, Slough, Berkshire SL1 3WE, UK
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Received: 12 July 2018 / Revised: 29 August 2018 / Accepted: 30 August 2018 / Published: 31 August 2018
(This article belongs to the Special Issue Bispecific Antibodies-Opportunities and Challenges)
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Abstract

Bispecific antibodies represent an emerging class of antibody drugs that are commonly generated by fusion of Fv or scFv antigen binding domains to IgG or Fab scaffolds. Fv- or scFv-mediated multimerisation of bispecific antibodies via promiscuous vH-vL pairing can result in sub-optimal monomer levels during expression, and hence, undesirable therapeutic protein yields. We investigate the contribution of disulphide stabilised Fv and scFv to Fab-Fv and Fab-scFv multimerisation. We show that monomer levels of isolated Fv/scFv cannot always be used to predict monomer levels of Fab-linked Fv/scFv, and that Fab-scFv monomer levels are greater than the equivalent Fab-Fv. Through grafting bispecifics with framework/CDR-‘swapped’ Fv and scFv, we show that monomer levels of disulphide stabilised Fab-Fv and Fab-scFv can be improved by Fv framework ‘swapping’. The Fab-Fv and Fab-scFv can be considered representative of the significant number of bispecific antibody formats containing appended Fv/scFv, as we also used Fv framework ‘swapping’ to increase the monomer level of an IgG-scFv bispecific antibody. This research may, therefore, be useful for maximising the monomeric yield of numerous pharmaceutically-relevant bispecific formats in pre-clinical development. View Full-Text
Keywords: bispecific antibody; disulphide stabilised Fv; disulphide stabilised single chain Fv; monomer; thermal stability bispecific antibody; disulphide stabilised Fv; disulphide stabilised single chain Fv; monomer; thermal stability
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Bhatta, P.; Humphreys, D.P. Relative Contribution of Framework and CDR Regions in Antibody Variable Domains to Multimerisation of Fv- and scFv-Containing Bispecific Antibodies. Antibodies 2018, 7, 35.

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