Identification and Affinity-Quantification of ß-Amyloid and α-Synuclein Polypeptides Using On-Line SAW-Biosensor-Mass Spectrometry

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Journal of The American Society for Mass Spectrometry. 2014, 25(8), pp. 1472-1481. ISSN 1044-0305. eISSN 1879-1123. Available under: doi: 10.1007/s13361-014-0904-1
Zusammenfassung

Bioaffinity analysis using a variety of biosensors has become an established tool for detection and quantification of biomolecular interactions. Biosensors, however, are generally limited by the lack of chemical structure information of affinity-bound ligands. On-line bioaffinity-mass spectrometry using a surface-acoustic wave biosensor (SAW-MS) is a new combination providing the simultaneous affinity detection, quantification, and mass spectrometric structural characterization of ligands. We describe here an on-line SAW-MS combination for direct identification and affinity determination, using a new interface for MS of the affinity-isolated ligand eluate. Key element of the SAW-MS combination is a microfluidic interface that integrates affinity-isolation on a gold chip, in-situ sample concentration, and desalting with a microcolumn for MS of the ligand eluate from the biosensor. Suitable MS- acquisition software has been developed that provides coupling of the SAW-MS interface to a Bruker Daltonics ion trap-MS, FTICR-MS, and Waters Synapt-QTOF- MS systems. Applications are presented for mass spectrometric identifications and affinity (KD) determinations of the neurodegenerative polypeptides, ß-amyloid (Aß), and pathophysiological and physiological synucleins (α- and ß-synucleins), two key polypeptide systems for Alzheimer’s disease and Parkinson’s disease, respectively. Moreover, first in vivo applications of αSyn polypeptides from brain homogenate show the feasibility of on-line affinity-MS to the direct analysis of biological material. These results demonstrate on-line SAW-bioaffinity-MS as a powerful tool for structural and quantitative analysis of biopolymer interactions.

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540 Chemie
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Analytical Chemistry, Biotechnology, Organic Chemistry, Proteomics, Bioinformatics
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ISO 690SLAMNOIU, Stefan, Camelia VLAD, Mihaela STUMBAUM, Adrian MOISE, Kathrin LINDNER, Nicole ENGEL, Mar VILANOVA, Mireia DIAZ, Christiaan KARREMAN, Marcel LEIST, Thomas CIOSSEK, Bastian HENGERER, Marta VILASECA, Michael PRZYBYLSKI, 2014. Identification and Affinity-Quantification of ß-Amyloid and α-Synuclein Polypeptides Using On-Line SAW-Biosensor-Mass Spectrometry. In: Journal of The American Society for Mass Spectrometry. 2014, 25(8), pp. 1472-1481. ISSN 1044-0305. eISSN 1879-1123. Available under: doi: 10.1007/s13361-014-0904-1
BibTex
@article{Slamnoiu2014Ident-29741,
  year={2014},
  doi={10.1007/s13361-014-0904-1},
  title={Identification and Affinity-Quantification of ß-Amyloid and α-Synuclein Polypeptides Using On-Line SAW-Biosensor-Mass Spectrometry},
  number={8},
  volume={25},
  issn={1044-0305},
  journal={Journal of The American Society for Mass Spectrometry},
  pages={1472--1481},
  author={Slamnoiu, Stefan and Vlad, Camelia and Stumbaum, Mihaela and Moise, Adrian and Lindner, Kathrin and Engel, Nicole and Vilanova, Mar and Diaz, Mireia and Karreman, Christiaan and Leist, Marcel and Ciossek, Thomas and Hengerer, Bastian and Vilaseca, Marta and Przybylski, Michael}
}
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