A modular IgG-scFv bispecific antibody topology
Open Access
- 17 December 2009
- journal article
- research article
- Published by Oxford University Press (OUP) in Protein Engineering, Design and Selection
- Vol. 23 (4), 221-228
- https://doi.org/10.1093/protein/gzp077
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.Keywords
This publication has 50 references indexed in Scilit:
- Improved Therapeutic Results by Pretargeted Radioimmunotherapy of Non–Hodgkin's Lymphoma with a New Recombinant, Trivalent, Anti-CD20, Bispecific AntibodyCancer Research, 2008
- Quantitative Spatiotemporal Analysis of Antibody Fragment Diffusion and Endocytic Consumption in Tumor SpheroidsCancer Research, 2008
- Antibody tumor penetration: Transport opposed by systemic and antigen-mediated clearanceAdvanced Drug Delivery Reviews, 2008
- Properties, production, and applications of camelid single-domain antibody fragmentsApplied Microbiology and Biotechnology, 2007
- Stably tethered multifunctional structures of defined composition made by the dock and lock method for use in cancer targetingProceedings of the National Academy of Sciences, 2006
- Recombinant approaches to IgG-like bispecific antibodiesActa Pharmacologica Sinica, 2005
- A Fully Human Recombinant IgG-like Bispecific Antibody to Both the Epidermal Growth Factor Receptor and the Insulin-like Growth Factor Receptor for Enhanced Antitumor ActivityJournal of Biological Chemistry, 2005
- Anti-HLA-DR/anti-DOTA Diabody Construction in a Modular Gene Design Platform: Bispecific Antibodies for Pretargeted RadioimmunotherapyCancer Biotherapy & Radiopharmaceuticals, 2001
- Stability engineering of antibody single-chain Fv fragmentsJournal of Molecular Biology, 2001
- Design and production of novel tetravalent bispecific antibodiesNature Biotechnology, 1997