Accurate Correction of the "bulk Response" in Surface Plasmon Resonance Sensing Provides New Insights on Interactions Involving Lysozyme and Poly(ethylene glycol)
Artikel i vetenskaplig tidskrift, 2022

Surface plasmon resonance is a very well-established surface sensitive technique for label-free analysis of biomolecular interactions, generating thousands of publications each year. An inconvenient effect that complicates interpretation of SPR results is the "bulk response"from molecules in solution, which generate signals without really binding to the surface. Here we present a physical model for determining the bulk response contribution and verify its accuracy. Our method does not require a reference channel or a separate surface region. We show that proper subtraction of the bulk response reveals an interaction between poly(ethylene glycol) brushes and the protein lysozyme at physiological conditions. Importantly, we also show that the bulk response correction method implemented in commercial instruments is not generally accurate. Using our method, the equilibrium affinity between polymer and protein is determined to be KD = 200 μM. One reason for the weak affinity is that the interaction is relatively short-lived (1/koff < 30 s). Furthermore, we show that the bulk response correction also reveals the dynamics of self-interactions between lysozyme molecules on surfaces. Besides providing new insights on important biomolecular interactions, our method can be widely applied to improve the accuracy of SPR data generated by instruments worldwide.

surface plasmon resonance

poly(ethylene glycol)

kinetics

quantification

lysozyme

Författare

Justas Svirelis

Chalmers, Kemi och kemiteknik, Tillämpad kemi

John Andersson

Chalmers, Kemi och kemiteknik, Tillämpad kemi

Anna Stradner

Lunds universitet

Andreas Dahlin

Chalmers, Kemi och kemiteknik, Tillämpad kemi

ACS Sensors

23793694 (eISSN)

Vol. In Press

Ämneskategorier

Fysikalisk kemi

Biofysik

Teoretisk kemi

DOI

10.1021/acssensors.2c00273

Mer information

Senast uppdaterat

2022-04-29