2. Ligand Immobilization
Amine coupling is the most widely used ligand immobilization strategy. In this strategy a surface carboxyl group is converted to a NHS ester by injecting a mix of EDC/NHS or Sulfo-NHS (e.g. 40 mg/mL EDC, 10 mg/mL NHS). The Protein (ligand) is then injected over the surface. A primary amine group on the protein displaces the NHS group and an amide bond is formed, covalently linking the protein to the surface. Unreacted NHS esters are then blocked by injecting 1 M ethanolamine pH 8.5. Many well-characterized alternative immobilization strategies are also available.
Alternate Strategies technical bulletin
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