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

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

SUBMITTER: Adams R 

PROVIDER: S-EPMC10213618 | biostudies-literature | 2023

REPOSITORIES: biostudies-literature