Simple and Rapid Procedure for “Sandwich” Enzyme ImmunoAssay Development Using Biotin Labeled Microwells (AvidPlate-Biotin)

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Abstract
We have developed a rapid and simple protocol for the development of sandwich enzyme immunoassays utilizing a modified microwell plate carrying biotin on its surface, Aviplate-Biotin. A single polyclonal antibody is conjugated with either a reporter enzyme or with biotin. These conjugates are mixed with antigen to form a ternary complex containing both enzyme and biotin tagged antibodies; this complex is then specifically captured by Aviplate-Biotin that has been treated with avidin. Once optimized the assay can be completed in slightly over 2 hours using conventional enzyme immunoassay instrumentation.

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