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TitlePeptide-conjugation induced conformational changes in human IgG1 observed by optimized negative-staining and individual-particle electron tomography.
Journal, issue, pagesSci Rep, Vol. 3, Page 1089, Year 2013
Publish dateJan 21, 2013
AuthorsHuimin Tong / Lei Zhang / Allan Kaspar / Matthew J Rames / Liqing Huang / Gary Woodnutt / Gang Ren /
PubMed AbstractPeptides 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 ...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.
External linksSci Rep / PubMed:23346347 / PubMed Central
MethodsEM (tomography)
Resolution16.5 - 16.6 Å
Structure data

EMDB-2285:
3D map of a peptide conjugated antibody particle by individual-particle electron tomography (IPET) and optimized negative-staining.
Method: EM (tomography) / Resolution: 16.5 Å

EMDB-2286:
3D map of another peptide conjugated antibody particle by individual-particle electron tomography (IPET) and optimized negative-staining.
Method: EM (tomography) / Resolution: 16.6 Å

Source
  • Homo sapiens (human)

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