Peptide-Conjugation Induced Conformational Changes in Human IgG1 Observed by Optimized Negative-Staining and Individual-Particle Electron Tomography
Open Access
- 21 January 2013
- journal article
- research article
- Published by Springer Science and Business Media LLC in Scientific Reports
- Vol. 3 (1), srep01089
- https://doi.org/10.1038/srep01089
Abstract
Peptides show much promise as potent and selective drug candidates. Fusing peptides to a scaffold monoclonal antibody produces a conjugated antibody which has the advantages of peptide activity yet also has the pharmacokinetics determined by the scaffold antibody. However, the conjugated antibody often has poor binding affinity to antigens that may be related to unknown structural changes. The study of the conformational change is difficult by conventional techniques because structural fluctuation under equilibrium results in multiple structures co-existing. Here, we employed our two recently developed electron microscopy (EM) techniques: optimized negative-staining (OpNS) EM and individual-particle electron tomography (IPET). Two-dimensional (2D) image analyses and three-dimensional (3D) maps have shown that the domains of antibodies present an elongated peptide-conjugated conformational change, suggesting that our EM techniques may be novel tools to monitor the structural conformation changes in heterogeneous and dynamic macromolecules, such as drug delivery vehicles after pharmacological synthesis and development.This publication has 40 references indexed in Scilit:
- Antitumor Efficacy of a Thrombospondin 1 Mimetic CovX-BodyTranslational Oncology, 2011
- Peptide-Based Drug Design: Here and NowMethods in molecular biology (Clifton, N.J.), 2008
- Chemically programmed antibodies: Endothelin receptor targeting CovX-Bodies™Bioorganic & Medicinal Chemistry Letters, 2007
- Monoclonal Antibodies Specific for the Empty Conformation of HLA-DR1 Reveal Aspects of the Conformational Change Associated with Peptide BindingOnline Journal of Public Health Informatics, 2004
- Conformational diversity and protein evolution – a 60-year-old hypothesis revisitedTrends in Biochemical Sciences, 2003
- Antibody Multispecificity Mediated by Conformational DiversityScience, 2003
- Identification of the platelet ADP receptor targeted by antithrombotic drugsNature, 2001
- Apoptosis and cancer drug targetingJCI Insight, 1999
- Long-circulating (sterically stabilized) liposomes for targeted drug deliveryTrends in Pharmacological Sciences, 1994
- Anti-transferrin receptor antibody and antibody-drug conjugates cross the blood-brain barrier.Proceedings of the National Academy of Sciences of the United States of America, 1991