title: Use of porous membranes modified with polyelectrolyte multilayers as substrates for protein arrays with low nonspecific adsorption.
abstract: Coating of substrates with polyelectrolyte multilayers terminated with poly(acrylic acid) (PAA) followed by activation of the free -COOH groups of PAA provides a surface that readily reacts with amine groups to allow covalent immobilization of antibodies. The use of this procedure to prepare arrays of antibodies in porous alumina supports facilitates construction of a flow-through system for analysis of fluorescently labeled antigens. Detection limits in the analysis of Cy5-labeled IgG are 0.02 ng/mL because of the high surface area of the alumina membrane, and the minimal diameter of the substrate pores results in binding limited by kinetics, not mass transport. Moreover, PAA-terminated films resist nonspecific protein adsorption, so blocking of antibody arrays with bovine serum albumin is not necessary. These microarrays are capable of effective analysis in 10% fetal bovine serum.
authors.affiliation: Department of Chemistry, Michigan State University, East Lansing, Michigan 48824, USA.
authors.forename: Jinhua
authors.initials: J
authors.lastname: Dai
authors.forename: Gregory L
authors.initials: GL
authors.lastname: Baker
authors.forename: Merlin L
authors.initials: ML
authors.lastname: Bruening
mesh_terms: D000180:Acrylic Resins / Q000737:chemistry; D000327:Adsorption; D000537:Aluminum Oxide / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D000937:Antigen-Antibody Reactions; D001672:Biocompatible Materials / Q000737:chemistry*; D002417:Cattle; D004573:Electrolytes / Q000737:chemistry* / Q000145:classification / Q000494:pharmacology; D005456:Fluorescent Dyes; D007118:Immunoassay; D007074:Immunoglobulin G / Q000737:chemistry*; D008567:Membranes, Artificial; D051379:Mice; D016062:Porosity; D040081:Protein Array Analysis; D011506:Proteins / Q000737:chemistry*; D011817:Rabbits; D051381:Rats; D012710:Serum Albumin, Bovine; D013499:Surface Properties
chemical_list: D000180:Acrylic Resins; D001672:Biocompatible Materials; D004573:Electrolytes; D005456:Fluorescent Dyes; D007074:Immunoglobulin G; D008567:Membranes, Artificial; D011506:Proteins; D012710:Serum Albumin, Bovine; C006903:carbopol 940; D000537:Aluminum Oxide