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Checkerboard immunoblotting recognizes twenty epitopes among the B subunit proteins of the cholera enterotoxin family
Author(s) -
Qu ZhenHai,
Finkelstein Richard A.
Publication year - 1993
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.11501401143
Subject(s) - epitope , enterotoxin , cholera toxin , monoclonal antibody , protein subunit , linear epitope , epitope mapping , conformational epitope , microbiology and biotechnology , escherichia coli , vibrio cholerae , toxin , ganglioside , biology , chemistry , fusion protein , antibody , biochemistry , bacteria , recombinant dna , gene , genetics
Checkerboard immunoblotting (CBIB) was used to analyze the reactions of a series of monoclonal antibodies with proteins of the cholera enterotoxin (CT) family, including heat labile enterotoxins (LTs) produced by diarrheagenic strains of Escherichia coli and genetically engineered chimeric proteins in which single amino acids of the CT‐B subunit protein or human (H) LT‐B sub‐unit protein were substituted for corresponding residues in porcine (P) LT‐B. The result indicated that there were at least twenty different patterns of reactivity suggesting that there are at least twenty recognizable epitopes among the proteins studied. An epitope which includes Ala 46 appears to be particularly important. This epitope is common to CT and H‐LT but not P‐LT, and the epitope is not blocked by the G ml ganglioside. Human convalescent sera react with this epitope. CBIB is a versatile technique for epitope analysis.