Location: Produce Safety and Microbiology ResearchTitle: Bioconjugation of antibodies to horseradish peroxidase (hrp)
Submitted to: Methods in Molecular Biology
Publication Type: Book / Chapter
Publication Acceptance Date: 4/4/2015
Publication Date: 7/14/2015
Citation: Hnasko, R.M. 2015. Bioconjugation of antibodies to horseradish peroxidase (hrp).In: Hnasko, R., editor. ELISA: Methods and protocols. Methods in Molecular Biology. 1st edition. New York, NY: Humana Press. p. 43-50. doi: 10.1007/978-1-4939-2742-5.
Interpretive Summary: This manuscript provides a detailed methodology for the chemical activation of antibodies to reporter molecules for their use in the detection of specific targets used in diagnostic assays.
Technical 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.