title: Bioconjugation of Antibodies to Horseradish Peroxidase (HRP).
abstract: The bioconjugation of an antibody to an enzymatic reporter such as horseradish peroxidase (HRP) affords an effective mechanism by which immunoassay detection of a target antigen can be achieved. The use of heterobifunctional cross-linkers to covalently link antibodies to HRP provides a simple and convenient means to maintain antibody affinity while imparting a functional reporter used for antigen detection. In this chapter, we describe a process by which Sulfo-SMCC is used to generate a stable maleimide-activated HRP that is reactive with sulfhydryl groups generated in antibodies by SATA-mediated thiolation.
authors.affiliation: Produce Safety and Microbiology Unit (PSM), 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, Robert.hnasko@ars.usda.gov.
authors.forename: Robert M
authors.initials: RM
authors.lastname: Hnasko
mesh_terms: D000906:Antibodies / Q000737:chemistry*; D001426:Bacterial Proteins / Q000737:chemistry; D002846:Chromatography, Affinity / Q000379:methods; D002853:Chromatography, Liquid / Q000379:methods; D003432:Cross-Linking Reagents / Q000737:chemistry*; D006735:Horseradish Peroxidase / Q000737:chemistry*; D006801:Humans; D055767:Immobilized Proteins / Q000737:chemistry; D018796:Immunoconjugates / Q000737:chemistry* / Q000302:isolation & purification; D008301:Maleimides / Q000737:chemistry; D011485:Protein Binding; D013388:Succinimides / Q000737:chemistry; D013438:Sulfhydryl Compounds / Q000737:chemistry; D013440:Sulfides / Q000737:chemistry
chemical_list: D000906:Antibodies; D001426:Bacterial Proteins; D003432:Cross-Linking Reagents; C034284:IgG Fc-binding protein, Streptococcus; D055767:Immobilized Proteins; D018796:Immunoconjugates; D008301:Maleimides; D013388:Succinimides; D013438:Sulfhydryl Compounds; D013440:Sulfides; C071675:sulfosuccinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate; C039011:N-hydroxysuccinimide S-acetylthioacetate; D006735:Horseradish Peroxidase