Ambient DESI and LESA-MS analysis of proteins adsorbed to a biomaterial surface using in-situ surface tryptic digestion

The detection and identification of proteins adsorbed onto biomaterial surfaces under ambient conditions has significant experimental advantages but has proven to be difficult to achieve with conventional measuring technologies. In this study, we present an adaptation of desorption electrospray ioni...

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Main Authors: Rao, Wei, Celiz, Adam D., Scurr, David J., Alexander, Morgan R., Barrett, David A.
Format: Article
Published: Springer Verlag 2013
Subjects:
Online Access:https://eprints.nottingham.ac.uk/2588/
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author Rao, Wei
Celiz, Adam D.
Scurr, David J.
Alexander, Morgan R.
Barrett, David A.
author_facet Rao, Wei
Celiz, Adam D.
Scurr, David J.
Alexander, Morgan R.
Barrett, David A.
author_sort Rao, Wei
building Nottingham Research Data Repository
collection Online Access
description The detection and identification of proteins adsorbed onto biomaterial surfaces under ambient conditions has significant experimental advantages but has proven to be difficult to achieve with conventional measuring technologies. In this study, we present an adaptation of desorption electrospray ionization (DESI) and liquid extraction surface analysis (LESA) mass spectrometry (MS) coupled with in-situ surface tryptic digestion to identify protein species from a biomaterial surface. Cytochrome c, myoglobin, and BSA in a combination of single and mixture spots were printed in an array format onto Permanox slides, followed by in-situ surface digestion and detection via MS. Automated tandem MS performed on surface peptides was able to identify the proteins via MASCOT. Limits of detection were determined for DESI-MS and a comparison of DESI and LESA-MS peptide spectra characteristics and sensitivity was made. DESI-MS images of the arrays were produced and analyzed with imaging multivariate analysis to automatically separate peptide peaks for each of the proteins within a mixture into distinct components. This is the first time that DESI and LESA-MS have been used for the in-situ detection of surface digested proteins on biomaterial surfaces and presents a promising proof of concept for the use of ambient MS in the rapid and automated analysis of surface proteins.
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spelling nottingham-25882020-05-04T20:18:30Z https://eprints.nottingham.ac.uk/2588/ Ambient DESI and LESA-MS analysis of proteins adsorbed to a biomaterial surface using in-situ surface tryptic digestion Rao, Wei Celiz, Adam D. Scurr, David J. Alexander, Morgan R. Barrett, David A. The detection and identification of proteins adsorbed onto biomaterial surfaces under ambient conditions has significant experimental advantages but has proven to be difficult to achieve with conventional measuring technologies. In this study, we present an adaptation of desorption electrospray ionization (DESI) and liquid extraction surface analysis (LESA) mass spectrometry (MS) coupled with in-situ surface tryptic digestion to identify protein species from a biomaterial surface. Cytochrome c, myoglobin, and BSA in a combination of single and mixture spots were printed in an array format onto Permanox slides, followed by in-situ surface digestion and detection via MS. Automated tandem MS performed on surface peptides was able to identify the proteins via MASCOT. Limits of detection were determined for DESI-MS and a comparison of DESI and LESA-MS peptide spectra characteristics and sensitivity was made. DESI-MS images of the arrays were produced and analyzed with imaging multivariate analysis to automatically separate peptide peaks for each of the proteins within a mixture into distinct components. This is the first time that DESI and LESA-MS have been used for the in-situ detection of surface digested proteins on biomaterial surfaces and presents a promising proof of concept for the use of ambient MS in the rapid and automated analysis of surface proteins. Springer Verlag 2013-12 Article PeerReviewed Rao, Wei, Celiz, Adam D., Scurr, David J., Alexander, Morgan R. and Barrett, David A. (2013) Ambient DESI and LESA-MS analysis of proteins adsorbed to a biomaterial surface using in-situ surface tryptic digestion. Journal of The American Society for Mass Spectrometry, 24 (12). pp. 1927-1936. ISSN 1044-0305 LESA-MS ; DESI-MS ; Ambient MS Protein arrays ; In-situ digestion ; MS imaging ; MCR analysis http://link.springer.com/article/10.1007%2Fs13361-013-0737-3 doi:10.1007/s13361-013-0737-3 doi:10.1007/s13361-013-0737-3
spellingShingle LESA-MS ; DESI-MS ; Ambient MS Protein arrays ; In-situ digestion ; MS imaging ; MCR analysis
Rao, Wei
Celiz, Adam D.
Scurr, David J.
Alexander, Morgan R.
Barrett, David A.
Ambient DESI and LESA-MS analysis of proteins adsorbed to a biomaterial surface using in-situ surface tryptic digestion
title Ambient DESI and LESA-MS analysis of proteins adsorbed to a biomaterial surface using in-situ surface tryptic digestion
title_full Ambient DESI and LESA-MS analysis of proteins adsorbed to a biomaterial surface using in-situ surface tryptic digestion
title_fullStr Ambient DESI and LESA-MS analysis of proteins adsorbed to a biomaterial surface using in-situ surface tryptic digestion
title_full_unstemmed Ambient DESI and LESA-MS analysis of proteins adsorbed to a biomaterial surface using in-situ surface tryptic digestion
title_short Ambient DESI and LESA-MS analysis of proteins adsorbed to a biomaterial surface using in-situ surface tryptic digestion
title_sort ambient desi and lesa-ms analysis of proteins adsorbed to a biomaterial surface using in-situ surface tryptic digestion
topic LESA-MS ; DESI-MS ; Ambient MS Protein arrays ; In-situ digestion ; MS imaging ; MCR analysis
url https://eprints.nottingham.ac.uk/2588/
https://eprints.nottingham.ac.uk/2588/
https://eprints.nottingham.ac.uk/2588/