Amine coupling through EDC/NHS: a practical approach

Methods Mol Biol. 2010;627:55-73. doi: 10.1007/978-1-60761-670-2_3.


Surface plasmon resonance (SPR) is one of the leading tools in biomedical research. The challenge in its use is the controlled positioning of one of the components of an interaction on a carefully designed surface. Many attempts in interaction analysis fail due to the non-functional or unsuccessful immobilization of a reactant onto the complex matrix of that surface. The most common technique for linking ligands covalently to a hydrophilic solid surface is amine coupling via reactive esters. In this chapter detailed methods and problem discussions will be given to assist in fast decision analysis to optimize immobilization and regeneration. Topics in focus are different coupling techniques for small and large molecules, streptavidin-biotin sandwich immobilization, and optimizing regeneration conditions.

MeSH terms

  • Aldehydes / chemistry
  • Amination
  • Amines / chemistry*
  • Biotin / chemistry
  • Carbodiimides / chemistry*
  • Immobilized Proteins / analysis
  • Immobilized Proteins / chemistry
  • Oxidation-Reduction
  • Streptavidin / chemistry
  • Succinimides / chemistry*
  • Surface Plasmon Resonance / methods*
  • Surface Properties


  • 1-ethyl-3-(3-(diethylamino)propyl)carbodiimide
  • Aldehydes
  • Amines
  • Carbodiimides
  • Immobilized Proteins
  • Succinimides
  • Biotin
  • Streptavidin
  • N-hydroxysuccinimide