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

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2010
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Drăguşanu, Mihaela
Tu, Tingting
Dragusanu, Mihaela
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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 â-amyloid (Aâ) epitope peptides bound to anti-Aâ 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-á-synuclein human á-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.

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ISO 690DRĂGUŞANU, Mihaela, Brînduşa-Alina PETRE, Stefan SLAMNOIU, Camelia VLAD, Tingting TU, Michael PRZYBYLSKI, Mihaela DRAGUSANU, 2010. On-line bioaffinity-electrospray mass spectrometry for simultaneous detection, identification, and quantification of protein-ligand interactions. In: Journal of The American Society for Mass Spectrometry. 2010, 21(10), pp. 1643-1648. ISSN 1044-0305. eISSN 1879-1123. Available under: doi: 10.1016/j.jasms.2010.06.011
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@article{Dragusanu2010Onlin-1091,
  year={2010},
  doi={10.1016/j.jasms.2010.06.011},
  title={On-line bioaffinity-electrospray mass spectrometry for simultaneous detection, identification, and quantification of protein-ligand interactions},
  number={10},
  volume={21},
  issn={1044-0305},
  journal={Journal of The American Society for Mass Spectrometry},
  pages={1643--1648},
  author={Drăguşanu, Mihaela and Petre, Brînduşa-Alina and Slamnoiu, Stefan and Vlad, Camelia and Tu, Tingting and Przybylski, Michael and Dragusanu, Mihaela}
}
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