Surface Plasmon Resonance Analysis of Heparin-Binding Angiogenic Growth Factors

Methods Mol Biol. 2016:1464:73-84. doi: 10.1007/978-1-4939-3999-2_7.

Abstract

Surface plasmon resonance (SPR) is an optical technique to evaluate biomolecular interactions. Briefly, SPR measures the capacity of two molecules to bind each other by detecting reflected light from a prism-gold film interface. One of the two putative interactants (called ligand) is chemically immobilized onto the gold film. When the sensor is exposed to a sample containing the second interactant (called analyte), its binding to the immobilized ligand causes a change of the refractive index of the material above the gold surface that is monitored as a real-time graph of the response units against time, producing a real-time graph called sensorgram. SPR has become a golden standard technology for label-free, real-time interaction analysis in basic research and drug discovery in a wide array of biomedical areas, including oncology and virology [1, 2]. Here we describe the exploitation of SPR for the study of the capacity of the pro-oncogenic, pro-angiogenic HIV-1 p17 matrix protein [3, 4] to bind to heparin, a structural analog of heparan sulfate proteoglycans (HSPGs) receptors, and for the identification of novel HSPGs-antagonists to be used as anti-p17 drugs.

Keywords: Affinity calculation; Biosensor; Heparin; Protein binding; Surface plasmon resonance.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biosensing Techniques
  • HIV Antigens / metabolism*
  • HIV-1 / metabolism
  • Heparin / metabolism*
  • Protein Binding
  • Surface Plasmon Resonance / methods*
  • gag Gene Products, Human Immunodeficiency Virus / metabolism*

Substances

  • HIV Antigens
  • gag Gene Products, Human Immunodeficiency Virus
  • p17 protein, Human Immunodeficiency Virus Type 1
  • Heparin