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Combination of biosensor and mass spectrometry

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  • Date: - -
  • Venue: Lecture theatre 5, Dipartimento di Scienze del Farmaco, 12 via Taramelli, Pavia
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On Monday the 18th of July Michael Przybylski from the Biopolymer Analysis and Biomedical Mass Spectrometry Unit in Rüsselsheim will give a seminar on Combination of biosensor and mass spectrometry. The seminar will be held in lecture theatre 5 (formerly lecture theatre B) at the Dipartiment of Scienze del Farmaco. The poster of the lecture can be downloaded here All members of the Department are invited to attend.

Abstract. Abstract. Bioaffinity analysis using biosensors has become an established technique for detection and quantification of biomolecular interactions. However, a principal limitation of biosensors is their lack of providing chemical structure information of affinity-bound ligands. Proteolytic excision/extraction (Protex-MS), hydrogen-deuterium exchange (HDX-MS) of peptide backbone hydrogens, and Fast- Photochemical Oxidation (FPOP) are major techniques for mass spectrometry based elucidation of protein- ligand interactions, but none of these tools alone provide quantitative affinity data. Using a surface plasmon resonance (SPR) biosensor, we have developed a continuous online biosensor-MS combination with electrospray ionization mass spectrometry that enables the simultaneous affinity isolation, structure identification and affinity quantification of biopolymer ligands from a protein- ligand complex immobilized on a gold chip. Key tool of the online biosensor-MS epitope analyzer is a new integrated, automated interface that provides sample concentration and in-situ desalting for the direct MS analysis of the ligand eluate [1]. ESI-MS systems from all major manufacturers can be coupled, using a newly developed software. The broad application potential of the online-SPR-MS epitope analyzer will be shown by recent studies of an unusual mixed-disulfide antibody epitope of the rheumatoic target protein, HLA-B27; and the interaction site identification of chaperone complexes of lysosomal enzymes [2, 3]. Interaction epitopes as diverse as antigen-antibody and lectin- carbohydrate complexes [4], and affinity binding constants (KD) from milli- to nanomolar ranges can be directly analysed. Applications amenable with the online-SPR-MS epitope analyzer include affinity-based biomarker identification, identification of protein and peptide epitopes, precise antibody affinity determinations, and direct label-free antigen quantification.

[1] Lupu, L. et al., (2016) Anal. Bioanal. Chem., in press; and Przybylski, M. et al., (2016) Eur. Pat. Appl.
[2] Iurascu, M. et al. (2015) J. Am. Soc. Mass Spectrom.
[3] Moise A., et al. (2016) J. Am. Soc. Mass Spectrom.,
[4] Moise, A., et al., (2011) J. Am. Chem. Soc. 133, 14844-14847.

Biography. Biography. Michael Przybylski studied chemistry and completed his PhD dissertation at the University of Mainz. After being research associate in Bioorganic Chemistry at Mainz, he spent two years as a Visiting Scientist at the National Cancer Institute, NIH/USA. He was Associate Professor in Organic and Biopolymer Chemistry at the University of Mainz, and was appointed in 1989 to the Professorship and Head of Analytical Chemistry at the University of Konstanz, where he is the Director of the Laboratory of Analytical Chemistry and Biopolymer Structure Analysis. Since then his laboratory has made numerous developments and applications of biopolymer mass spectrometry methods, combined with protein‐ and peptide‐chemical methods, tertiary structure analysis by protein‐chemical modification and mass spectrometry, and mass spectrometric determination of biopolymer recognition structures. His laboratory has invented proteolytic‐excision mass spectrometry for the elucidation of protein‐lig and interaction structures, and peptide/protein epitopes, and has elucidated the structure of several membrane proteins, such as Lung Surfactant protein‐C. The laboratory’s current research is focused on applications of mass spectrometry and peptide biochemistry to structure and mechanism of neurodegenerative proteins; pathophysiological protein modification; vaccine chemistry; structure and epitope analysis of therapeutic antibodies; affinity‐mass spectrometry of biopolymer interaction epitopes; mass spectrometric elucidation of oligomerisation/truncation pathways and structures of “misfolding”/aggregating proteins in neurodegenerative diseases. Michael Przybylski has been awarded the St Denis Prize in Cancer Research; several international Guest Professorships; the Life Science Prize of the German Society for Mass Spectrometry; an honorary doctoral degree of the University of Iasi, RO; He is currently Guest Professor of the Chinese Academy of Sciences, Changchun, and of Jilin University,China; he has been recently appointed Adjunct Professor of Chemistry of Indiana University, Bloomington, Indiana, USA. He has been member of Scientific Committees of several International Conferences, has been Editor and is Editorial Board member of several International Journals in the field of biopolymer chemistry and mass spectrometry, and has been Chairman of the German Society for Mass Spectrometry. He has been coordinator of two previous FP5 and FP6 EU projects. He has served as scientific advisor, and board member of several international Research Institutions in mass spectrometry and proteomics, and is member of the Graduate School “Chemical Biology”, and board member of the “Zukunftskolleg”, two recently appointed Excellence Centers of the University of Konstanz. He has ca. 300 publications in International Journals; 6 monographs/reviews; ca. 25 patents; ca. 100 invited lectures.

Image. Zhang Y et al. Interface for Online Coupling of Surface Plasmon Resonance to Direct Analysis in Real Time Mass Spectrometry. Anal Chem 87:6505 (2015)








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