Tandem Mass Spectrometry-Based Amyloid Typing Using Manual Microdissection and Open-Source Data Processing.
Amyloid
Crux
Liquid chromatography
Open source
Tandem mass spectrometry
Journal
American journal of clinical pathology
ISSN: 1943-7722
Titre abrégé: Am J Clin Pathol
Pays: England
ID NLM: 0370470
Informations de publication
Date de publication:
04 05 2022
04 05 2022
Historique:
received:
21
07
2021
accepted:
20
09
2021
entrez:
5
5
2022
pubmed:
6
5
2022
medline:
10
5
2022
Statut:
ppublish
Résumé
Standard implementations of amyloid typing by liquid chromatography-tandem mass spectrometry use capabilities unavailable to most clinical laboratories. To improve accessibility of this testing, we explored easier approaches to tissue sampling and data processing. We validated a typing method using manual sampling in place of laser microdissection, pairing the technique with a semiquantitative measure of sampling adequacy. In addition, we created an open-source data processing workflow (Crux Pipeline) for clinical users. Cases of amyloidosis spanning the major types were distinguishable with 100% specificity using measurements of individual amyloidogenic proteins or in combination with the ratio of λ and κ constant regions. Crux Pipeline allowed for rapid, batched data processing, integrating the steps of peptide identification, statistical confidence estimation, and label-free protein quantification. Accurate mass spectrometry-based amyloid typing is possible without laser microdissection. To facilitate entry into solid tissue proteomics, newcomers can leverage manual sampling approaches in combination with Crux Pipeline and related tools.
Identifiants
pubmed: 35512256
pii: 6433227
doi: 10.1093/ajcp/aqab185
pmc: PMC9071319
doi:
Substances chimiques
Amyloid
0
Amyloidogenic Proteins
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
748-757Subventions
Organisme : NIGMS NIH HHS
ID : R01 GM121818
Pays : United States
Informations de copyright
© American Society for Clinical Pathology, 2021. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.
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