Methods for Epitope Mapping

Bioassay-based or enzyme-linked immunosorbent assay (ELISA) methods have been applied in epitope mapping [18, 19]. In this approach, different antigens are coated separately on the surface in different weUs of the analytical plate followed by incubation with a specific antibody that is linked to an enzyme. When the enzyme s substrate is added to the solution, a subsequent reaction can produce a detectable signal. This approach, however, is mostly limited to the characterization of linear epitopes. ELISA can also be applied to multisite binding analysis, where one antibody is attached to the surface, the antigen is bound, and the ability of the second antibody to bind to the attached antibody-antigen 

Immunochemical analysis - Yeast display -Bioassay (ELISA) 

SPR measurements. The information such as the specificity and binding afBnity of antibody-antigen can be obtained in a single SPR expaiment. 

In general, immunochemical analyses of antibody-antigen interactions can provide both qualitative and quantitative information. Howevo-, these approaches lack the ability to provide information regarding the detailed locations of epitopes on antigens. In order to obtain such information, one must conduct structural analysis on antibody-antigen complexes. 

MS-based strategies based on cross-linking and protein footprinting provide alternative approaches for epitope mapping [31]. Although these methods cannot reveal three-dimensional structures of antibody-antigen complexes, the information on the location of both linear and conformational epitopes can be obtained. In addition, the sample amount required for MS measurements is small (femtomole to picomole), and there is no known upper mass limit when a proteolytic step is included. 

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Morris, G. E. (1996) Choosing a method for epitope mapping, in Epitope Mapping Protocols (G.E. Morris, ed.), Humana Press. Totowa, NJ, pp. 1-9. 

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