Production of antibodies against degradative neoepitopes in aggrecan

Methods Mol Med. 2004:100:237-50. doi: 10.1385/1-59259-810-2:237.

Abstract

The use of synthetic peptides to generate rabbit polyclonal anticatabolic neoepitope antibodies that can be used to study the presence of defined proteolytic cleavage sites in aggrecan is described. Principles of peptide design and methods for preparation and characterization of ovalbumin conjugates are presented along with approaches for the characterization and affinity purification of the resulting antisera. Limitations associated with the use of antipeptide antibodies to study authentic protein neoepitopes are discussed.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aggrecans
  • Amino Acid Sequence
  • Animals
  • Antibodies / analysis
  • Antibodies / immunology
  • Epitopes / chemistry
  • Epitopes / immunology*
  • Extracellular Matrix Proteins / chemistry
  • Extracellular Matrix Proteins / immunology*
  • Immune Sera / biosynthesis*
  • Immune Sera / isolation & purification
  • Lectins, C-Type
  • Molecular Sequence Data
  • Peptides / chemistry
  • Peptides / immunology*
  • Proteoglycans / chemistry
  • Proteoglycans / immunology*
  • Rabbits
  • Succinimides / chemistry

Substances

  • Aggrecans
  • Antibodies
  • Epitopes
  • Extracellular Matrix Proteins
  • Immune Sera
  • Lectins, C-Type
  • Peptides
  • Proteoglycans
  • Succinimides
  • N-succinimidyl bromoacetate