A Modular IgG-scFv Bispecific Antibody Topology

Overview

Journal Protein Eng Des Sel

Publisher Oxford University Press

Specialties Biochemistry
Biotechnology

Date 2009 Dec 19

PMID 20019028

Citations 63

Authors

Kelly Davis Orcutt

Margaret E Ackerman

Maryelise Cieslewicz

Emmanuel Quiroz

Adrian L Slusarczyk

John V Frangioni

K Dane Wittrup

Affiliations

Soon will be listed here.

Abstract

Here we present a bispecific antibody (bsAb) format in which a disulfide-stabilized scFv is fused to the C-terminus of the light chain of an IgG to create an IgG-scFv bifunctional antibody. When expressed in mammalian cells and purified by one-step protein A chromatography, the bsAb retains parental affinities of each binding domain, exhibits IgG-like stability and demonstrates in vivo IgG-like tumor targeting and blood clearance. The extension of the C-terminus of the light chain of an IgG with an scFv or even a smaller peptide does appear to disrupt disulfide bond formation between the light and heavy chains; however, this does not appear to affect binding, stability or in vivo properties of the IgG. Thus, we demonstrate here that the light chain of an IgG can be extended with an scFv without affecting IgG function and stability. This format serves as a standardized platform for the construction of functional bsAbs.

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