title: Effect of the Protein Corona on Antibody-Antigen Binding in Nanoparticle Sandwich Immunoassays.
abstract: We investigated the effect of the protein corona on the function of nanoparticle (NP) antibody (Ab) conjugates in dipstick sandwich immunoassays. Ab specific for Zika virus nonstructural protein 1 (NS1) were conjugated to gold NPs, and another anti-NS1 Ab was immobilized onto the nitrocellulose membrane. Sandwich immunoassay formation was influenced by whether the strip was run in corona forming conditions, i.e., in human serum. Strips run in buffer or pure solutions of bovine serum albumin exhibited false positives, but those run in human serum did not. Serum pretreatment of the nitrocellulose also eliminated false positives. Corona formation around the NP-Ab in serum was faster than the immunoassay time scale. Langmuir binding analysis determined how the immobilized Ab affinity for the NP-Ab/NS1 was impacted by corona formation conditions, quantified as an effective dissociation constant, KDeff. Results show that corona formation mediates the specificity and sensitivity of the antibody-antigen interaction of Zika biomarkers in immunoassays, and plays a critical but beneficial role.
authors.forename: Helena
authors.initials: H
authors.lastname: de Puig
authors.forename: Irene
authors.initials: I
authors.lastname: Bosch
authors.affiliation: Department of Chemical Engineering, Institut Quimic de Sarria, Universitat Ramon Llull , Via Augusta 390, 08017 Barcelona, Spain.
authors.forename: Marc
authors.initials: M
authors.lastname: Carré-Camps
authors.affiliation: Department of Engineering, University of Massachusetts Boston , Boston, Massachusetts 02125, United States.
authors.forename: Kimberly
authors.identifier: 0000-0002-4839-3179
authors.initials: K
authors.lastname: Hamad-Schifferli
mesh_terms: D000937:Antigen-Antibody Reactions; D015415:Biomarkers; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D053758:Nanoparticles / Q000737:chemistry*; D000066970:Protein Corona / Q000378:metabolism*; D020349:Surface Plasmon Resonance; D000071244:Zika Virus / Q000276:immunology
chemical_list: D015415:Biomarkers; D000066970:Protein Corona