EFab domain substitution as a solution to the light-chain pairing problem of bispecific antibodies

MAbs
Heather A CookeMartin Preyer

Abstract

Bispecific antibody therapeutics can expand the functionality of a conventional monoclonal antibody drug because they can bind multiple antigens. However, their great potential is counterbalanced by the challenges faced in their production. The classic asymmetric bispecific containing an Fc requires the expression of four unique chains - two light chains and two heavy chains; each light chain must pair with its correct heavy chain, which then must heterodimerize to form the full bispecific. The light-chain pairing problem has several solutions, some of which require engineering and optimization for each bispecific pair. Here, we introduce a technology called EFab Domain Substitution, which replaces the Cε2 of IgE for one of the CL/CH1 domains into one arm of an asymmetric bispecific to encourage the correct pairing of the light chains. EFab Domain Substitution provides very robust correct pairing while maintaining antibody function and is effective for many variable domains. We report its effect on the biophysical properties of an antibody and the crystal structure of the EFab domain substituted into the adalimumab Fab (PDB ID 6CR1).

References

Dec 31, 1987·Biochemical and Biophysical Research Communications·Y KuwanaY Kurosawa
Oct 21, 1988·Science·R E BirdM Whitlow
Oct 6, 1983·Nature·C Milstein, A C Cuello
Jan 3, 2001·Acta Crystallographica. Section D, Biological Crystallography·M D WinnG N Murshudov
Feb 13, 2001·The Journal of Immunology : Official Journal of the American Association of Immunologists·N F LandolfiN Tsurushita
Jun 18, 2002·Nature Immunology·Tommy WanBrian J Sutton
Dec 2, 2004·Acta Crystallographica. Section D, Biological Crystallography·Paul Emsley, Kevin Cowtan
Dec 2, 2004·Acta Crystallographica. Section D, Biological Crystallography·Alexei A VaginGarib N Murshudov
Feb 25, 2006·Journal of Molecular Biology·Robyn L StanfieldBernhard Rupp
Feb 4, 2010·Acta Crystallographica. Section D, Biological Crystallography·Paul D AdamsPeter H Zwart
Apr 26, 2011·Nature Structural & Molecular Biology·Mary D HoldomBrian J Sutton
Jun 22, 2011·Proceedings of the National Academy of Sciences of the United States of America·Wolfgang SchaeferChristian Klein
Oct 5, 2013·Molecular Cancer Therapeutics·Oliver SeifertRoland E Kontermann
Jan 28, 2014·Nature Biotechnology·Steven M LewisStephen J Demarest
Feb 1, 2015·Molecular Immunology·Christoph SpiessPaul J Carter
Jan 1, 1997·Methods in Enzymology·Zbyszek Otwinowski, Wladek Minor
Jan 7, 2017·Proceedings of the National Academy of Sciences of the United States of America·Patrick KoenigGermaine Fuh
Jan 11, 2017·MAbs·Ulrich Brinkmann, Roland E Kontermann
Jan 18, 2017·Proceedings of the National Academy of Sciences of the United States of America·Tushar JainK Dane Wittrup

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Citations

Jun 9, 2019·Nature Reviews. Drug Discovery·Aran F LabrijnPaul W H I Parren

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Methods Mentioned

BETA
glycosylation
biosensors
bio-layer interferometry
differential scanning calorimetry
X-ray
surface plasmon resonance
PCR
transfection
size-exclusion chromatography
chip

Software Mentioned

Bio
MOLREP
HKL2000
MaxLynx
Coot
REFMAC
Phenix
MaxEnt
Rad Gel Doc EZ

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