Peptide-Conjugation Induced Conformational Changes in Human IgG1 Observed by Optimized Negative-Staining and Individual-Particle Electron Tomography

Tong, Huimin; Zhang, Lei; Kaspar, Allan; Rames, Matthew J.; Huang, Liqing; Woodnutt, Gary; Ren, Gang

doi:10.1038/srep01089

Peptide-Conjugation Induced Conformational Changes in Human IgG1 Observed by Optimized Negative-Staining and Individual-Particle Electron Tomography

Journal Article · Mon Jan 21 00:00:00 EST 2013 · Scientific Reports

DOI:https://doi.org/10.1038/srep01089· OSTI ID:1624583

Tong, Huimin ^[1]; Zhang, Lei ^[2]; Kaspar, Allan ^[3]; Rames, Matthew J. ^[2]; Huang, Liqing ^[4]; Woodnutt, Gary ^[3]; Ren, Gang ^[2]

Xi'an Jiaotong Univ., Shaanxi (China). School of Science. MOE Key Lab. for Non-equilibrium Synthesis and Modulation of Condensed Matter; Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Molecular Foundry; DOE/OSTI
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Molecular Foundry
Pfizer Inc, CovX Research LLC, San Diego, CA (United States)
Xi'an Jiaotong Univ., Shaanxi (China). School of Science. MOE Key Lab. for Non-equilibrium Synthesis and Modulation of Condensed Matter

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.

View Accepted Manuscript (DOE)

Research Organization:: Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Basic Energy Sciences (BES). Chemical Sciences, Geosciences & Biosciences Division

Grant/Contract Number:: AC02-05CH11231

OSTI ID:: 1624583

Journal Information:: Scientific Reports, Journal Name: Scientific Reports Journal Issue: 1 Vol. 3; ISSN 2045-2322

Publisher:: Nature Publishing GroupCopyright Statement

Country of Publication:: United States

Language:: English

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Peptide-Conjugation Induced Conformational Changes in Human IgG1 Observed by Optimized Negative-Staining and Individual-Particle Electron Tomography

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