Tandem Mass Spectrometry-Based Amyloid Typing Using Manual Microdissection and Open-Source Data Processing.
Objectives: 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.Methods: We validated a...
| Publicado en: | American Journal of Clinical Pathology Vol. 157; no. 5; pp. 748 - 758 |
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| Autores principales: | , , , , , , , , , , , |
| Formato: | Journal Article |
| Publicado: |
Oxford University Press / USA
May2022
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| Acceso en línea: | Ver este registro en EBSCOhost |
| fields | @attributes: recordID: 1 pdfLink: plink: https://search.ebscohost.com/login.aspx?direct=true&db=ccm&AN=156677904&site=ehost-live header: @attributes: shortDbName: ccm uiTerm: 156677904 longDbName: CINAHL Complete uiTag: AN controlInfo: bkinfo: dissinfo: jinfo: jid: 00029173 2RW jtl: American Journal of Clinical Pathology issn: 00029173 maglogo: N pubinfo: dt: May2022 vid: 157 iid: 5 pid: 10398 pub: Oxford University Press / USA artinfo: ui: 156677904 156677904 NLM35512256 10.1093/ajcp/aqab185 NLM35512256 156677904 ppf: 748 ppct: 10 formats: tig: atl: Tandem Mass Spectrometry-Based Amyloid Typing Using Manual Microdissection and Open-Source Data Processing. aug: au: Phipps, William S Smith, Kelly D Yang, Han-Yin Henderson, Clark M Pflaum, Hannah Lerch, Melissa L Fondrie, William E Emrick, Michelle A Wu, Christine C MacCoss, Michael J Noble, William S Hoofnagle, Andrew N affil: Department of Laboratory Medicine and Pathology , Seattle, WA , USA sug: subj: Amyloidosis Diagnosis Mass Spectrometry Methods Proteins Analysis Specimen Handling Proteins Clinical Assessment Tools Scales ab: Objectives: 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.Methods: 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.Results: 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.Conclusions: 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. pubtype: Academic Journal doctype: Journal Article ougenre: Article language: English refInfo: holdings: @attributes: islocal: N |
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