A Protein Suspension-Trapping Sample Preparation for Tear Proteomics by Liquid Chromatography-Tandem Mass Spectrometry.
J Vis Exp
; (202)2023 Dec 01.
Article
in En
| MEDLINE
| ID: mdl-38108381
ABSTRACT
Tear fluid is one of the easily accessible biofluids that can be collected non-invasively. Tear proteomics has the potential to discover biomarkers for several ocular diseases and conditions. The suspension trapping column has been reported to be an efficient and user-friendly sample preparation workflow for the broad application of downstream proteomic analysis. Yet, this strategy has not been well-studied in the analysis of human tear proteome. The present protocol describes an integrated workflow from clinical human tear samples to purified peptides for non-invasive tear protein biomarker research using mass spectrometry, which provides insights into disease biomarkers and monitoring when combined with bioinformatics analysis. A protein suspension trapping sample preparation was applied and demonstrated the discovery of tear proteome with fast, reproducible, and user-friendly procedures, as a universal, optimized sample preparation for human tear fluid analysis. In particular, the suspension trapping procedure outperformed in-solution sample preparation in terms of peptide recovery, protein identification, and shorter sample preparation time.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Proteome
/
Lacerations
Limits:
Humans
Language:
En
Journal:
J Vis Exp
/
J. vis. exp
/
Journal of visualized experiments
Year:
2023
Document type:
Article
Country of publication: