Proteomic analysis of human cataract aqueous humour: Comparison of one-dimensional gel LCMS with two-dimensional LCMS of unlabelled and iTRAQ®-labelled specimens.
Author(s): Bennett KL, Funk M, Tschernutter M, Breitwieser FP, Planyavsky M, Ubaida Mohien C, Müller A, Trajanoski Z, Colinge J, Superti-Furga G, Schmidt-Erfurth U
Publication: J Proteomics, 2011, Vol. 74, Page 151-66
PubMed ID: 20940065 PubMed Review Paper? No
Purpose of Paper
Conclusion of Paper
Studies
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Study Purpose
The purpose of this study was to compare the proteomes of aqueous humor identified using three strategies for dissociation. Aqueous humor specimens from patients undergoing cataracts surgery were stored with protease inhibitor at 4 degrees C, for less than 4 h, prior to centrifugation and storage of the supernatant at -80 degrees C.
Summary of Findings:
While 156 protein groups were identified by one-dimensional gel collision-induced dissociation (1D-G-CID), 117 and 77 protein groups were identified by two-dimensional shotgun collision-induced dissociation (2D-SG-CID) and two-dimensional shotgun high-energy collision-induced dissociation (2D-SG-HCD), respectively. Of the 198 total protein groups identified, 58 were identified by all three methods, and 36 protein groups were identified by two of three methods. iTRAQ labeling of 2D-SG-HCD specimens showed that 88% of proteins were expressed similarly (<2 fold difference) between specimens.
Biospecimens
Preservative Types
- Frozen
Diagnoses:
- Cataracts
Platform:
Analyte Technology Platform Protein LC-MS or LC-MS/MS Protein Fluorometry Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) LC-MS or LC-MS/MS Specific Detection method iTRAQ label
None
LC-MS or LC-MS/MS Specific Technology platform One-dimensional gel collision-induced dissociation
Two-dimensional shotgun collision-induced dissociation
Two-dimensional shotgun high-energy collision-induced dissociation