TY - JOUR
T1 - Peptide sequencing directly on solid surfaces using MALDI mass spectrometry
AU - Zhao, Zhan-Gong
AU - Cordovez, Lalaine Anne
AU - Johnston, Stephen
AU - Woodbury, Neal
N1 - Funding Information:
We thank Geneva Nguyen and Alesha Robertson for their technical assistance, and Dr. Douglas Daniel for design of ammonia gas chamber. This work was supported by a contract to S.A.J. from the Department of Homeland Security (HSHQDC-15-C-B0008), the NSF INSPIRE grant: MCB-1243082 to N.W., the SRA grant from HealthTell, Inc. to Z.G.Z. and the Center for Innovations in Medicine Peptide Array Core.
Publisher Copyright:
© 2017 The Author(s).
PY - 2017/12/1
Y1 - 2017/12/1
N2 - There are an increasing variety of applications in which peptides are both synthesized and used attached to solid surfaces. This has created a need for high throughput sequence analysis directly on surfaces. However, common sequencing approaches that can be adapted to surface bound peptides lack the throughput often needed in library-based applications. Here we describe a simple approach for sequence analysis directly on solid surfaces that is both high speed and high throughput, utilizing equipment available in most protein analysis facilities. In this approach, surface bound peptides, selectively labeled at their N-termini with a positive charge-bearing group, are subjected to controlled degradation in ammonia gas, resulting in a set of fragments differing by a single amino acid that remain spatially confined on the surface they were bound to. These fragments can then be analyzed by MALDI mass spectrometry, and the peptide sequences read directly from the resulting spectra.
AB - There are an increasing variety of applications in which peptides are both synthesized and used attached to solid surfaces. This has created a need for high throughput sequence analysis directly on surfaces. However, common sequencing approaches that can be adapted to surface bound peptides lack the throughput often needed in library-based applications. Here we describe a simple approach for sequence analysis directly on solid surfaces that is both high speed and high throughput, utilizing equipment available in most protein analysis facilities. In this approach, surface bound peptides, selectively labeled at their N-termini with a positive charge-bearing group, are subjected to controlled degradation in ammonia gas, resulting in a set of fragments differing by a single amino acid that remain spatially confined on the surface they were bound to. These fragments can then be analyzed by MALDI mass spectrometry, and the peptide sequences read directly from the resulting spectra.
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U2 - 10.1038/s41598-017-18105-3
DO - 10.1038/s41598-017-18105-3
M3 - Article
C2 - 29259225
AN - SCOPUS:85038612501
SN - 2045-2322
VL - 7
JO - Scientific reports
JF - Scientific reports
IS - 1
M1 - 17811
ER -