title: Antibody orientation enhanced by selective polymer-protein noncovalent interactions.
abstract: A unique interaction has been found between protein G' (a truncated recombinant bacterial "alphabet" protein which aligns by noncovalent attachment to the antibody stem) and poly(methyl methacrylate), a thermoplastic polymer substrate, which can be easily fabricated using high-rate processes. Significantly improved orientation efficiency with traditional passive adsorption for this system (termed ALYGNSA) has been achieved as compared to the same assay performed on a polystyrene substrate with protein G'. Results were consistent with an average alignment of 80% of the human immunoglobulin G capture antibody which translated into a 30% to 50% improved alignment over an array of industry standards tested. Laser scanning confocal microscopy confirmed the immunological results. Studies of additional poly(methyl methacrylate) polymer derivatives and protein biolinker (A and AG) combinations have been conducted and have revealed different degrees of antibody alignment. These findings may lead to additional novel noncovalent methods of antibody orientation and greater sensitivity in immunological assays.
authors.affiliation: Department of Chemistry, University of Massachusetts, Lowell, MA, 01854, USA.
authors.forename: Lisa-Jo A
authors.initials: LJ
authors.lastname: Clarizia
authors.forename: Davin
authors.initials: D
authors.lastname: Sok
authors.forename: Ming
authors.initials: M
authors.lastname: Wei
authors.forename: Joey
authors.initials: J
authors.lastname: Mead
authors.forename: Carol
authors.initials: C
authors.lastname: Barry
authors.forename: Melisenda J
authors.initials: MJ
authors.lastname: McDonald
mesh_terms: D005453:Fluorescence; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism*; D007834:Lasers; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D019904:Polymethyl Methacrylate / Q000737:chemistry / Q000378:metabolism*; D011494:Protein Kinases / Q000378:metabolism
chemical_list: C029152:G-substrate; D007074:Immunoglobulin G; D009419:Nerve Tissue Proteins; D019904:Polymethyl Methacrylate; D011494:Protein Kinases