title: Real-time analysis of antibody interactions with whole enveloped human cytomegalovirus using surface plasmon resonance.
abstract: Human cytomegalovirus (CMV) is a large enveloped virus that encodes multiple glycoproteins required for virus-cell binding and fusion. To assess the binding properties of antibodies with target glycoprotein in a natural context of infection, we investigated the feasibility of using the surface plasmon resonance (SPR) technique for studying the direct binding of antibodies with CMV virions. Direct immobilization of whole virions to sensor surface and a surface regeneration procedure allowed for quantitative and reproducible measurements of binding affinity and binding kinetics of antibody-whole virion interactions. The conformational and functional integrity of viral particles was not compromised by the regeneration condition as evaluated with antibodies recognizing conformational epitopes and by electron microscopy. Binding of an irrelevant antibody was not observed, indicating the high specificity of the method. A panel of anti-gB antibodies was measured and the binding affinities correlated fairly well with those determined by ELISA. These data demonstrated that the interaction of anti-gB antibody with whole virion of large enveloped CMV can be quantitatively studied using SPR. This method has been successfully applied for screening and selection of anti-CMV antibodies and can be potentially extended to study antibody-glycoprotein interactions of other related herpesviruses.
authors.affiliation: Infectious Diseases, Novartis Institutes for BioMedical Research, Inc., Cambridge, MA 02139, USA.
authors.forename: Gregg
authors.initials: G
authors.lastname: Chenail
authors.forename: Nathan E
authors.initials: NE
authors.lastname: Brown
authors.forename: Ashley
authors.initials: A
authors.lastname: Shea
authors.forename: Adam L
authors.initials: AL
authors.lastname: Feire
authors.forename: Gejing
authors.initials: G
authors.lastname: Deng
mesh_terms: D000914:Antibodies, Viral / Q000276:immunology*; D000918:Antibody Specificity / Q000276:immunology; D003587:Cytomegalovirus / Q000276:immunology* / Q000648:ultrastructure; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007700:Kinetics; D015203:Reproducibility of Results; D020349:Surface Plasmon Resonance / Q000379:methods*; D013499:Surface Properties; D014759:Viral Envelope Proteins / Q000276:immunology*; D014771:Virion / Q000648:ultrastructure
chemical_list: D000914:Antibodies, Viral; D014759:Viral Envelope Proteins