Epitope mapping of monoclonal antibodies to keratin 19 using keratin fragments, synthetic peptides and phage peptide libraries

Eur J Biochem. 1995 Jul 15;231(2):475-85. doi: 10.1111/j.1432-1033.1995.tb20721.x.

Abstract

To generate tools for monitoring processing and folding in keratin intermediate filaments, a group of monoclonal antibodies reacting with the intermediate filament protein keratin 19 were studied using different approaches to define the structure and localization of their epitopes. The binding pattern to bacterially expressed human keratin 19 fragments allowed the definition of minimal amino acid sequences required for antibody binding. The screening of overlapping 15-residue peptides confirmed and further specified the epitope locations for a subset of the tested antibodies. In addition, the epitope of an antibody with apparent species-restricted specificity (LE64) was revealed by isolating and characterizing a full-length keratin 19 clone from a PtK2 cDNA library. Taken together with species cross-reactivity of individual antibodies and sequence information obtained by probing a phage display library, specific amino acid residues could be highlighted as likely to be involved in the antibody binding.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Antibodies, Monoclonal / immunology*
  • Bacteriophages / genetics
  • Base Sequence
  • Binding Sites, Antibody
  • Epitope Mapping*
  • Epitopes / immunology
  • Gene Library
  • Humans
  • Immunoblotting
  • Keratins / chemistry
  • Keratins / genetics
  • Keratins / immunology*
  • Molecular Sequence Data
  • Peptide Fragments / chemistry
  • Peptide Fragments / genetics
  • Peptide Fragments / immunology
  • Peptides / chemistry
  • Peptides / genetics
  • Peptides / immunology
  • Protein Structure, Secondary
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / immunology
  • Sequence Alignment
  • Sequence Homology, Nucleic Acid
  • Species Specificity

Substances

  • Antibodies, Monoclonal
  • Epitopes
  • Peptide Fragments
  • Peptides
  • Recombinant Fusion Proteins
  • Keratins

Associated data

  • GENBANK/X82579