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Positive reactions on Western blots do not necessarily indicate the epitopes on antigens are continuous
Author(s) -
Zhou YiHua,
Chen Zhaochun,
Purcell Robert H,
Emerson Suzanne U
Publication year - 2007
Publication title -
immunology and cell biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.999
H-Index - 104
eISSN - 1440-1711
pISSN - 0818-9641
DOI - 10.1038/sj.icb.7100004
Subject(s) - epitope , blot , monoclonal antibody , antigen , antibody , microbiology and biotechnology , gel electrophoresis , epitope mapping , polyacrylamide gel electrophoresis , biology , chemistry , virology , enzyme , biochemistry , immunology , gene
Epitope mapping (identification of an antigenic site recognized by an antibody) is an important component of vaccine development and immunological assays. It is widely accepted that in Western blots, antibodies react exclusively with continuous epitopes: discontinuous epitopes are assumed to be irreversibly destroyed by electrophoresis under the denaturing conditions used for sodium dodecyl sulfate‐polyacrylamide gel electrophoresis. Here, we demonstrate that the epitopes recognized by four different monoclonal antibodies were identified as discontinuous epitopes when characterized by radioimmunoprecipitation assays and enzyme‐linked immunosorbent assays, yet each of these antibodies reacted with the corresponding antigen on Western blots. Reaction on Western blots may be due to epitope renaturation during or after the transfer of the protein to a membrane. Therefore, positive reactions on Western blots do not necessarily indicate that epitopes are continuous and this caveat should be kept in mind while characterizing them.