Serum albumin binding knob domains engineered within a VH framework III bispecific antibody format and as chimeric peptides

DOI: 10.3389/fimmu.2023.1170357

Authors: Ralph Adams (UCB Biopharma UK) , Callum Joyce (UCB Biopharma UK) , Mikhail Kuravskiy (UCB Biopharma UK) , Katriona Harrison (The University of Sydney) , Zainab Ahdash (UCB Biopharma UK) , Matthew Balmforth (UCB Biopharma UK) , Kelda Chia (UCB Biopharma UK) , Cinzia Marceddu (UCB Biopharma UK) , Matthew Coates (UCB Biopharma UK) , James Snowden (UCB Biopharma UK) , Emmanuel Goursaud (UCB Biopharma SA) , Karelle Ménochet (UCB Biopharma UK) , Jean Van Den Elsen (University of Bath) , Richard J. Payne (The University of Sydney) , Alastair D. G. Lawson (UCB Biopharma UK) , Anthony Scott-Tucker (UCB Biopharma UK) , Alex Macpherson (UCB Biopharma UK)
Co-authored by industrial partner: Yes

Type: Journal Paper
Journal: Frontiers In Immunology , VOL 14

State: Published (Approved)
Published: May 2023
Diamond Proposal Number(s): 29404 , 5956

Abstract: Background: Serum albumin binding is an established mechanism to extend the serum half-life of antibody fragments and peptides. The cysteine rich knob domains, isolated from bovine antibody ultralong CDRH3, are the smallest single chain antibody fragments described to date and versatile tools for protein engineering. Methods: Here, we used phage display of bovine immune material to derive knob domains against human and rodent serum albumins. These were used to engineer bispecific Fab fragments, by using the framework III loop as a site for knob domain insertion. Results: By this route, neutralisation of the canonical antigen (TNFα) was retained but extended pharmacokinetics in-vivo were achieved through albumin binding. Structural characterisation revealed correct folding of the knob domain and identified broadly common but non-cross-reactive epitopes. Additionally, we show that these albumin binding knob domains can be chemically synthesised to achieve dual IL-17A neutralisation and albumin binding in a single chemical entity. Conclusions: This study enables antibody and chemical engineering from bovine immune material, via an accessible discovery platform.

Journal Keywords: knob domain; bispecific; albumin; ultralong CDRH3; peptide

Subject Areas: Biology and Bio-materials, Chemistry, Medicine


Instruments: I02-Macromolecular Crystallography , I03-Macromolecular Crystallography

Added On: 07/06/2023 10:58

Documents:
fimmu-14-1170357.pdf

Discipline Tags:

Biochemistry Chemistry Structural biology Chemical Engineering Engineering & Technology Life Sciences & Biotech

Technical Tags:

Diffraction Macromolecular Crystallography (MX)