Epitope Mapping by Region-Specified PCR Mutagenesis
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This technique was developed for mapping of epitopes of monoclonal antibodies (MAb) at the amino acid sequence level; i.e., the identification of amino acids of an antigenic protein involved in specific interactions with each antibody. The epitope mapping includes:
| 1. |
Construction of a random base substitution DNA library for the gene encoding the antigenic protein in the λ-gt11 phage expression-vector;
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| 2. |
Tests for the crossreactivities of proteins expressed in plaques formed by phages in the library; and
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| 3. |
An analysis of DNA sequences of the mutant genes encoding proteins with altered crossreactivities.
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