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On-line bioaffinity-electrospray mass spectrometry for simultaneous detection, identification, and quantification of protein-ligand interactions

  • Mihaela Dragusanu
  • Brîndusa-Alina Petre
  • Stefan Slamnoiu
  • Camelia Vlad
  • Tingting Tu
  • Michael PrzybylskiEmail author
Focus: Affinity Mass Spectrometry

Abstract

We describe here an on-line combination of a surface acoustic wave (SAW) biosensor with electrospray ionization mass spectrometry (SAW-ESI-MS) that enables the direct detection, identification, and quantification of affinity-bound ligands from a protein-ligand complex on a biosensor chip. A trapping column was used between the SAW-biosensor and the electrospray mass spectrometer equipped with a micro-guard column, which provides simultaneous sample concentration and desalting for the mass spectrometric analysis of the dissociated ligand. First applications of the on-line SAW-ESI-MS combination include (1), differentiation of β-amyloid (Aβ) epitope peptides bound to anti-Aβ antibodies; (2), the identification of immobilized Substance P peptide-calmodulin complex; (3), identification and quantification of the interaction of 3-nitrotyrosine-modified peptides with nitrotyrosine-specific antibodies; and (4), identification of immobilized anti-α-synuclein-human α-synuclein complex. Quantitative determinations of protein-ligand complexes by SAW yielded dissociation constants (KD) from micro-to low nanomolar sample concentrations. The on-line bioaffinity-ESI-MS combination presented here is expected to enable broad bioanalytical application to the simultaneous, label-free determination and quantification of biopolymer-ligand interactions, as diverse as antigen-antibody and lectin-carbohydrate complexes.

Keywords

Surface Plasmon Resonance Surface Acoustic Wave Mass Spectrometric Analysis Isothermal Titration Calorimetry Trapping Column 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© American Society for Mass Spectrometry 2010

Authors and Affiliations

  • Mihaela Dragusanu
    • 1
  • Brîndusa-Alina Petre
    • 1
  • Stefan Slamnoiu
    • 1
  • Camelia Vlad
    • 1
  • Tingting Tu
    • 2
  • Michael Przybylski
    • 1
    Email author
  1. 1.Laboratory of Analytical Chemistry and Biopolymer Structure Analysis, Department of ChemistryUniversity of KonstanzKonstanzGermany
  2. 2.Department of ChemistryWashington UniversitySt. LouisUSA

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