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

  • Research Article
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Journal of The American Society for Mass Spectrometry

Abstract

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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Acknowledgments

This work was supported in part by grant KF2026662 from the Bundesministerium für Wirtschaft und Technologie, Berlin, Germany. Support by the European Union within the Marie Curie IRSES project Integrating High Performance Mass Spectrometry Tools with Applications in Life Sciences is gratefully acknowledged.

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Author notes

  1. Camelia Vlad & Mihaela Stumbaum

    Present address: SAW- Instruments, Schwertberger Strasse 16, 53177, Bonn, Germany

  2. Nicole Engel

    Present address: Department of Chemical Technology, Technical University of Vienna, 1060, Vienna, Austria

Authors and Affiliations

  1. Laboratory of Analytical Chemistry and Steinbeis Center for Biopolymer Structure Analysis, University of Konstanz, 78457, Konstanz, Germany

    Stefan Slamnoiu, Camelia Vlad, Mihaela Stumbaum, Adrian Moise, Kathrin Lindner, Nicole Engel & Michael Przybylski

  2. Department of Biology, University of Konstanz, 78457, Konstanz, Germany

    Christiaan Karreman & Marcel Leist

  3. CNS Research Division, Boehringer Ingelheim Pharma, Biberach, Germany

    Thomas Ciossek & Bastian Hengerer

  4. IRB Barcelona - Institute for Research in Biomedicine, Barcelona, Spain

    Mar Vilanova, Mireia Diaz & Marta Vilaseca

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  1. Stefan Slamnoiu
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Corresponding author

Correspondence to Michael Przybylski.

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Slamnoiu, S., Vlad, C., Stumbaum, M. et al. Identification and Affinity-Quantification of ß-Amyloid and α-Synuclein Polypeptides Using On-Line SAW-Biosensor-Mass Spectrometry. J. Am. Soc. Mass Spectrom. 25, 1472–1481 (2014). https://doi.org/10.1007/s13361-014-0904-1

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  • DOI: https://doi.org/10.1007/s13361-014-0904-1

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