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Peptide epitope binding and fluorescence quenching with the anti-mucin antibody C595
M J Scanlon1, M R Price, S J Tendler
1Department of Pharmaceutical Sciences, University of Nottingham, University Park, UK.
The Fab fragment of the anti-polymorphic epithelial mucin antibody C595 (IgG3, kappa) has been produced by digestion with papain. The purified antibody fragment retained its binding activity for the mucin in an indirect radioisotopic antiglobulin assay. The binding constant of the Fab fragment for the synthetic peptide (PDTRPAPGSTAPPAHGVTSA) corresponding to the 20-amino acid tandem repeat sequence found in the mucin protein core was determined to be 0.3 x 10(6) M-1 by measuring the capacity of the peptide to quench the fluorescence of the Fab fragment.
The Fab fragment of the anti-polymorphic epithelial mucin antibody C595 (IgG3, kappa) has been produced by digestion with papain. The purified antibody fragment retained its binding activity for the mucin in an indirect radioisotopic antiglobulin assay. The binding constant of the Fab fragment for the synthetic peptide (PDTRPAPGSTAPPAHGVTSA) corresponding to the 20-amino acid tandem repeat sequence found in the mucin protein core was determined to be 0.3 x 10(6) M-1 by measuring the capacity of the peptide to quench the fluorescence of the Fab fragment.