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Epitope mapping by region-specified PCR-mutagenesis

Tsutomu Mikawa1, Masayuki Lkeda, Takehiko Shibata

  • 1Biometal Science Laboratory, RIKEN SPring-8 Center, Hyogo, Japan.

Methods in Molecular Biology (Clifton, N.J.)
|April 21, 2009
PubMed

We will describe a procedure to map epitopes on a protein against monoclonal IgGs. In this procedure, we amplified and mutagenized the entire or a part of the protein. Then, a DNA region encoding the protein was cut out by a restriction enzyme and ligated into a lambda-gt11 expression vector to construct a library. Thus, the protein is expressed as a fusion protein with beta-galactosidase. Protein in plaques obtained by phages derived from the library were tested for cross-reactivities by means of immunoblotting experiments.

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