Fab-based bispecific antibody formats with robust biophysical properties and biological activity

MAbs
Xiufeng WuStephen J Demarest

Abstract

A myriad of innovative bispecific antibody (BsAb) platforms have been reported. Most require significant protein engineering to be viable from a development and manufacturing perspective. Single-chain variable fragments (scFvs) and diabodies that consist only of antibody variable domains have been used as building blocks for making BsAbs for decades. The drawback with Fv-only moieties is that they lack the native-like interactions with CH1/CL domains that make antibody Fab regions stable and soluble. Here, we utilize a redesigned Fab interface to explore 2 novel Fab-based BsAbs platforms. The redesigned Fab interface designs limit heavy and light chain mixing when 2 Fabs are co-expressed simultaneously, thus allowing the use of 2 different Fabs within a BsAb construct without the requirement of one or more scFvs. We describe the stability and activity of a HER2×HER2 IgG-Fab BsAb, and compare its biophysical and activity properties with those of an IgG-scFv that utilizes the variable domains of the same parental antibodies. We also generated an EGFR × CD3 tandem Fab protein with a similar format to a tandem scFv (otherwise known as a bispecific T cell engager or BiTE). We show that the Fab-based BsAbs have superior biophysical p...Continue Reading

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Citations

Aug 16, 2016·Current Opinion in Structural Biology·Mark L Chiu, Gary L Gilliland
Jan 11, 2017·MAbs·Ulrich Brinkmann, Roland E Kontermann
Jun 16, 2017·Journal of Chemical Theory and Computation·Bernhard KnappCharlotte M Deane
Jul 21, 2017·Protein Science : a Publication of the Protein Society·K J FroningS J Demarest
Dec 11, 2019·Antibodies·Mark L ChiuGary L Gilliland
Sep 7, 2019·Science Advances·Johan M S van der SchootFerenc A Scheeren
May 3, 2018·Protein Engineering, Design & Selection : PEDS·Xiufeng WuStephen J Demarest
Aug 23, 2018·BioDrugs : Clinical Immunotherapeutics, Biopharmaceuticals and Gene Therapy·Bushra Husain, Diego Ellerman
May 13, 2020·Nature Communications·Karen FroningStephen J Demarest
Jan 24, 2021·Proceedings of the National Academy of Sciences of the United States of America·Wentao ChenJames A Wells
Oct 16, 2020·International Journal of Molecular Sciences·Manali S SawantPeter M Tessier
Aug 24, 2017·Pharmacology & Therapeutics·Z Wu, N V Cheung
May 25, 2021·Frontiers in Immunology·Jiabing MaCheng Qian
Jul 14, 2021·Journal for Immunotherapy of Cancer·Mathilde PoussinStephen Demarest
Dec 1, 2017·Journal of the American Chemical Society·Juanjuan DuFeng Wang

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Methods Mentioned

BETA
circular
size
static
proteomic
transfection
static light scattering
surface plasmon resonance
chip
circular dichroism
FACS

Software Mentioned

Diva
Global3
GraphPad Prism

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