title: A Double-Sandwich ELISA for Identification of Monoclonal Antibodies Suitable for Sandwich Immunoassays.
abstract: The sandwich immunoassay (sELISA) is an invaluable technique for concentrating, detecting, and quantifying target antigens. The two critical components required are a capture antibody and a detection antibody, each binding a different epitope on the target antigen. The specific antibodies incorporated into the test define most of the performance parameters of any subsequent immunoassay regardless of the assay format: traditional ELISA, lateral-flow immunoassay, various bead-based assays, antibody-based biosensors, or the reporting label. Here we describe an approach for identifying monoclonal antibodies (mAbs) suitable for use as capture antibodies and detector antibodies in a sELISA targeting bacterial protein toxins. The approach was designed for early identification of monoclonal antibodies (mAbs), in the initial hybridoma screen.
authors.affiliation: Foodborne Toxin Detection and Prevention Research Unit (FTDP), Western Regional Research Center (WRRC), Pacific West Area (PWA), Agricultural Research Service (ARS), United States Department of Agriculture (USDA), 800 Buchanan St., Albany, CA, 94710, USA, Larry.stanker@ars.usda.gov.
authors.forename: Larry H
authors.initials: LH
authors.lastname: Stanker
authors.forename: Robert M
authors.initials: RM
authors.lastname: Hnasko
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification*; D019274:Botulinum Toxins, Type A / Q000302:isolation & purification*; D017077:Culture Media, Conditioned / Q000737:chemistry; D004797:Enzyme-Linked Immunosorbent Assay / Q000295:instrumentation / Q000379:methods*; D057166:High-Throughput Screening Assays; D006735:Horseradish Peroxidase / Q000737:chemistry*; D006801:Humans; D006825:Hybridomas / Q000378:metabolism; D018796:Immunoconjugates / Q000737:chemistry*; D007141:Immunoglobulin Fc Fragments / Q000737:chemistry*; D008027:Light; D049449:Luminescence; D011485:Protein Binding; D011817:Rabbits; D012680:Sensitivity and Specificity
chemical_list: D000911:Antibodies, Monoclonal; D017077:Culture Media, Conditioned; D018796:Immunoconjugates; D007141:Immunoglobulin Fc Fragments; D006735:Horseradish Peroxidase; D019274:Botulinum Toxins, Type A