Identification and affinity-quantification of ß-amyloid and α-synuclein polypeptides using on-line SAW-biosensor-mass spectrometry
| Autor: | Marta Vilaseca, Christiaan Karreman, Adrian Moise, Bastian Hengerer, Camelia Vlad, Michael Przybylski, Mihaela Stumbaum, Marcel Leist, Nicole Engel, Mireia Diaz, Kathrin Lindner, Stefan Slamnoiu, Thomas Ciossek, Mar Vilanova |
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| Rok vydání: | 2013 |
| Předmět: |
Chemical structure
Antibody Affinity Mice Transgenic Biosensing Techniques engineering.material Proteomics Mass spectrometry Mass Spectrometry Epitopes Structural Biology Animals Humans Amino Acid Sequence Spectroscopy Neurons Chromatography Amyloid beta-Peptides Fourier Analysis Chemistry Ligand Elution Antibodies Monoclonal Brain Cyclotrons Microfluidic Analytical Techniques Recombinant Proteins Molecular Weight Sound Amino Acid Substitution ddc:540 engineering Analytical Chemistry Biotechnology Organic Chemistry Proteomics Bioinformatics alpha-Synuclein Feasibility Studies Mutant Proteins Biopolymer Biosensor Quantitative analysis (chemistry) |
| Zdroj: | Journal of the American Society for Mass Spectrometry. 25(8) |
| ISSN: | 1879-1123 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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