Peptide-conjugation induced conformational changes in human IgG1 observed by optimized negative-staining and individual-particle electron tomography.
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.
MOE Key Laboratory for Non-equilibrium Synthesis and Modulation of Condensed Matter, School of Science, Xi'an Jiaotong University, Xi'an 710049, China.
Huimin
H
Tong
Lei
L
Zhang
Allan
A
Kaspar
Matthew J
MJ
Rames
Liqing
L
Huang
Gary
G
Woodnutt
Gang
G
Ren
D000906:Antibodies / Q000737:chemistry / Q000276:immunology; D000941:Antigens / Q000737:chemistry / Q000276:immunology; D055032:Electron Microscope Tomography / Q000379:methods; D006801:Humans; D018796:Immunoconjugates / Q000737:chemistry* / Q000276:immunology; D007074:Immunoglobulin G / Q000737:chemistry* / Q000276:immunology; D046911:Macromolecular Substances / Q000737:chemistry* / Q000276:immunology; D008854:Microscopy, Electron / Q000379:methods*; D008968:Molecular Conformation; D016624:Negative Staining / Q000379:methods*; D010455:Peptides / Q000737:chemistry* / Q000276:immunology
D000906:Antibodies; D000941:Antigens; D018796:Immunoconjugates; D007074:Immunoglobulin G; D046911:Macromolecular Substances; D010455:Peptides