Enzyme-Linked Immunosorbent Assay of Peptides
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Enzyme-linked immunosorbent assay (ELISA) is a member of the solid-phase immunoassay family that detects specific antigen-antibody
binding reactions (1
,2
). A great many variables of antigen or antibody presentation, treatment, and detection are considered when performing ELISA
(3
,4
). This chapter will describe the simplest form, a direct assay, in which the peptide antigen is immobilized on a polystyrene
carrier, and a specific primary antibody in solution (detector) is added that recognizes the plate-bound antigen. A secondary,
horseradish peroxidase (HRP)-coupled polyclonal antibody (reporter) binds to the primary antibody. The enzymatic activation
of hydrogen peroxide releases a chromophore from o
-phenylenediamine that is detected and quantified.