Surface Plasmon Resonance Method to Evaluate Anti-citrullinated Protein/Peptide Antibody Affinity to Citrullinated Peptides

Methods Mol Biol. 2015:1348:267-74. doi: 10.1007/978-1-4939-2999-3_23.

Abstract

Surface plasmon resonance (SPR) technique is extremely interesting in immunology because it has the potential to directly visualize biomolecular interactions in real-time monitoring antibody affinity, one of the parameters affecting pathogenicity in autoimmune diseases. Herein we describe the affinity evaluation of anti-citrullinated peptide antibodies (ACPA) to a peptide-based biosensor by SPR. The method describes the purification of ACPA isolated from rheumatoid arthritis (RA) patients using affinity columns, the strategy employed for the immobilization of citrullinated peptides onto a sensor chip, and the evaluation of the specific binding of purified ACPA to immobilized peptides.

Keywords: Antibody affinity; Biacore; Citrullinated peptides; Kinetic; Surface plasmon resonance (SPR).

MeSH terms

  • Antibodies / chemistry*
  • Antibodies / immunology*
  • Antibodies / isolation & purification
  • Antibody Affinity / immunology*
  • Chromatography, Affinity
  • Citrulline / chemistry*
  • Hydrogen-Ion Concentration
  • Kinetics
  • Peptides / chemistry*
  • Peptides / immunology*
  • Surface Plasmon Resonance / methods*

Substances

  • Antibodies
  • Peptides
  • Citrulline