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Regenerable Biosensors for Small-Molecule Kinetic Characterization Using SPR.

Anders Gunnarsson1, Christopher J Stubbs2, Philip B Rawlins2

  • 1Structure, Biophysics and Fragment-based Lead Generation, Discovery Sciences, R&D, AstraZeneca, Gothenburg, Sweden.

SLAS Discovery : Advancing Life Sciences R & D
|December 8, 2020
PubMed
Summary

New immobilization strategies for surface plasmon resonance (SPR) improve drug discovery throughput. These methods combine irreversible and reversible techniques, enabling kinetic characterization of challenging compounds and targets.

Keywords:
surface immobilizationsurface regeneration

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Area of Science:

  • Biochemistry
  • Biophysics
  • Drug Discovery

Background:

  • Characterizing compound-target interactions is crucial in small-molecule drug discovery.
  • Surface plasmon resonance (SPR) is a versatile technique for this, offering wide affinity range and kinetic analysis.
  • Traditional SPR immobilization methods (covalent, streptavidin-biotin) are irreversible, limiting throughput for unstable targets or slowly dissociating compounds. Reversible methods (His-tag-Ni-NTA) often suffer from baseline drift, hindering accurate kinetic quantification.

Purpose of the Study:

  • To investigate novel immobilization strategies for SPR that combine the stability of irreversible methods with the flexibility of reversible ones.
  • To address limitations in throughput and kinetic characterization of challenging analytes in SPR assays.
  • To expand the SPR immobilization toolbox for previously out-of-scope applications.

Main Methods:

  • Investigated three immobilization strategies: dual-His-tagged target protein, His-tagged streptavidin, and switchavidin.
  • Evaluated the advantages and limitations of each strategy for SPR applications.
  • Applied these strategies to kinetic characterization of kinase inhibitors and covalent inhibitors.

Main Results:

  • Developed and assessed three novel SPR immobilization techniques.
  • Demonstrated that these strategies can overcome limitations of traditional methods.
  • Achieved significantly increased throughput for kinetic characterization of potent kinase inhibitors and covalent inhibitors.

Conclusions:

  • The investigated immobilization strategies offer a balance between robustness and flexibility for SPR.
  • While a universal method is yet to be found, these approaches enhance the SPR toolbox.
  • These advancements enable broader applications, including high-throughput kinetic profiling of drug candidates.