Rapid Surface Shaving for Proteomic Identification of Novel Surface Antigens for Vaccine Development.

Luu, LDW; Lan, R

Rapid Surface Shaving for Proteomic Identification of Novel Surface Antigens for Vaccine Development.

Luu, LDW Lan, R

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Publisher:: Springer Nature
Publication Type:: Journal Article
Citation:: Methods Mol Biol, 2022, 2414, pp. 47-62
Issue Date:: 2022

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Field	Value	Language
dc.contributor.author	Luu, LDW
dc.contributor.author	Lan, R
dc.date.accessioned	2023-03-20T01:12:03Z
dc.date.available	2023-03-20T01:12:03Z
dc.date.issued	2022
dc.identifier.citation	Methods Mol Biol, 2022, 2414, pp. 47-62
dc.identifier.isbn	9781071618998
dc.identifier.issn	1064-3745
dc.identifier.issn	1940-6029
dc.identifier.uri	http://hdl.handle.net/10453/167682
dc.description.abstract	The bacterial cell surface (surfaceome) is the first site encountered by immune cells and is thus an important site for immune recognition. As such, the characterization of bacterial surface proteins can lead to the discovery of novel antigens for potential vaccine development. In this chapter, we describe a rapid 5-min surface shaving proteomics protocol where live bacterial cells are incubated with trypsin and surface peptides are "shaved" off. The shaved peptides are subsequently identified with liquid chromatography-tandem mass spectrometry (LC-MS/MS). Several checkpoints, including colony forming unit (CFU) counts, flow cytometry, and a false positive unshaved control, are introduced to ensure cell viability/membrane integrity are maintained and that proteins identified are true surface proteins. The protein topology of shaved peptides can be bioinformatically confirmed for surface location. Surface shaving facilitates identification of surface proteins expressed under different conditions, by different strains as well as highly abundant essential and immunogenic bacterial surface antigens for potential vaccine development.
dc.format	Print
dc.language	eng
dc.publisher	Springer Nature
dc.relation.ispartof	Methods Mol Biol
dc.relation.isbasedon	10.1007/978-1-0716-1900-1_4
dc.rights	info:eu-repo/semantics/closedAccess
dc.subject	0399 Other Chemical Sciences, 0601 Biochemistry and Cell Biology
dc.subject.classification	Developmental Biology
dc.subject.mesh	Antigens, Bacterial
dc.subject.mesh	Antigens, Surface
dc.subject.mesh	Bacteria
dc.subject.mesh	Bacterial Proteins
dc.subject.mesh	Chromatography, Liquid
dc.subject.mesh	Membrane Proteins
dc.subject.mesh	Peptides
dc.subject.mesh	Proteomics
dc.subject.mesh	Tandem Mass Spectrometry
dc.subject.mesh	Vaccines
dc.subject.mesh	Bacteria
dc.subject.mesh	Peptides
dc.subject.mesh	Bacterial Proteins
dc.subject.mesh	Membrane Proteins
dc.subject.mesh	Vaccines
dc.subject.mesh	Antigens, Bacterial
dc.subject.mesh	Antigens, Surface
dc.subject.mesh	Chromatography, Liquid
dc.subject.mesh	Proteomics
dc.subject.mesh	Tandem Mass Spectrometry
dc.subject.mesh	Antigens, Bacterial
dc.subject.mesh	Antigens, Surface
dc.subject.mesh	Bacteria
dc.subject.mesh	Bacterial Proteins
dc.subject.mesh	Chromatography, Liquid
dc.subject.mesh	Membrane Proteins
dc.subject.mesh	Peptides
dc.subject.mesh	Proteomics
dc.subject.mesh	Tandem Mass Spectrometry
dc.subject.mesh	Vaccines
dc.title	Rapid Surface Shaving for Proteomic Identification of Novel Surface Antigens for Vaccine Development.
dc.type	Journal Article
utslib.citation.volume	2414
utslib.location.activity	United States
utslib.for	0399 Other Chemical Sciences
utslib.for	0601 Biochemistry and Cell Biology
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Science
pubs.organisational-group	/University of Technology Sydney/Faculty of Science/School of Life Sciences
utslib.copyright.status	closed_access	*
dc.date.updated	2023-03-20T01:12:01Z
pubs.publication-status	Published
pubs.volume	2414

Abstract:

The bacterial cell surface (surfaceome) is the first site encountered by immune cells and is thus an important site for immune recognition. As such, the characterization of bacterial surface proteins can lead to the discovery of novel antigens for potential vaccine development. In this chapter, we describe a rapid 5-min surface shaving proteomics protocol where live bacterial cells are incubated with trypsin and surface peptides are "shaved" off. The shaved peptides are subsequently identified with liquid chromatography-tandem mass spectrometry (LC-MS/MS). Several checkpoints, including colony forming unit (CFU) counts, flow cytometry, and a false positive unshaved control, are introduced to ensure cell viability/membrane integrity are maintained and that proteins identified are true surface proteins. The protein topology of shaved peptides can be bioinformatically confirmed for surface location. Surface shaving facilitates identification of surface proteins expressed under different conditions, by different strains as well as highly abundant essential and immunogenic bacterial surface antigens for potential vaccine development.

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http://hdl.handle.net/10453/167682