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Using Surface Plasmon Resonance to Quantitatively Assess Lipid–Protein Interactions

  • Kathryn Del Vecchio
  • Robert V. StahelinEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1376)

Abstract

Surface Plasmon Resonance (SPR) is a quantitative, label-free method for determining molecular interactions in real time. The technology involves fixing a ligand onto a senor chip, measuring a baseline resonance angle, and flowing an analyte in bulk solution over the fixed ligand to measure the subsequent change in resonance angle. The mass of analyte bound to fixed ligand is directly proportional to the resonance angle change and the system is sensitive enough to detect as little as picomolar amounts of analyte in the bulk solution. SPR can be used to determine both the specificity of molecular interactions and the kinetics and affinity of an interaction. This technique has been especially useful in measuring the affinities of lipid-binding proteins to intact liposomes of varying lipid compositions.

Key words

Binding affinity Equilibrium binding Kinetics Lipid–protein interactions Surface plasmon resonance 

Notes

Acknowledgement

The NIH (AI081077) and NSF (1122068) have supported lipid–protein interaction work in the Stahelin lab using SPR. K.D. is funded by a CBBI NIH T32 Predoctoral fellowship (NIH T32GM075762).

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Department of Chemistry and BiochemistryUniversity of Notre DameNotre DameUSA
  2. 2.Department of Biochemistry and Molecular BiologyIndiana University School of Medicine-South BendSouth BendUSA

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