Technical advance: Surface plasmon resonance-based analysis of CXCL12 binding using immobilized lentiviral particles

J Leukoc Biol. 2011 Aug;90(2):399-408. doi: 10.1189/jlb.1010565. Epub 2011 May 18.


Use of SPR-based biosensors is an established method for measuring molecular interactions. Their application to the study of GPCRs is nonetheless limited to detergent-solubilized receptors that can then be reconstituted into a lipid environment. Using the chemokine receptor CXCR4 and its specific ligand CXCL12, we outline here a highly reproducible biosensor method based on receptor presentation on the surface of lentiviral particles; the approach is simple and does not require the use of antibodies to achieve correct receptor orientation on the sensorchip surface. We measured the kinetic parameters of CXCR4/CXCL12 binding in a single step and in real time and evaluated the effect of GAG presentation of chemokines on this interaction. The data indicate that at low concentrations, soluble heparin modulates CXCR4/CXCL12 interaction and at high concentrations, abrogates binding. These observations suggest that in addition to their known role in modulating local chemokine availability, GAG affect the receptor/ligand interaction, although their influence on affinity parameters is very limited. The method will also be useful for quantifying these biomarkers in biological fluids and for the development of high-throughput screening for their antagonists.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biosensing Techniques / methods*
  • Chemokine CXCL12 / analysis*
  • Chemokine CXCL12 / metabolism*
  • Glycosaminoglycans / pharmacology
  • Heparin / pharmacology
  • Humans
  • Immobilized Proteins
  • Kinetics
  • Lentivirus / metabolism
  • Protein Binding
  • Receptors, CXCR4 / metabolism
  • Surface Plasmon Resonance


  • Chemokine CXCL12
  • Glycosaminoglycans
  • Immobilized Proteins
  • Receptors, CXCR4
  • Heparin