Addressing Trypsin Bias in Large Scale (Phospho)proteome Analysis by Size Exclusion Chromatography and Secondary Digestion of Large Post-Trypsin Peptides

2011 ◽  
Vol 10 (2) ◽  
pp. 800-811 ◽  
Author(s):  
Bao Quoc Tran ◽  
Celine Hernandez ◽  
Patrice Waridel ◽  
Alexandra Potts ◽  
Jachen Barblan ◽  
...  
Keyword(s):  
Size Exclusion Chromatography ◽  
Large Scale ◽  
Proteome Analysis ◽  
Size Exclusion ◽  
Exclusion Chromatography

Large-scale Purification of a Lentiviral Vector by Size Exclusion Chromatography or Mustang Q Ion Exchange Capsule

2003 ◽  
Vol 2 (5) ◽  
pp. 89-95 ◽  
Author(s):  
Boro Dropulic ◽  
Vladimir Slepushkin ◽  
Nancy Chang ◽  
Yuxiang Gan ◽  
Bing Jiang ◽  
...  
Keyword(s):  
Ion Exchange ◽  
Size Exclusion Chromatography ◽  
Large Scale ◽  
Lentiviral Vector ◽  
Size Exclusion ◽  
Exclusion Chromatography

scholarly journals Mass-Spectrometry Based Proteome Comparison of Extracellular Vesicle Isolation Methods: Comparison of ME-kit, Size-Exclusion Chromatography, and High-Speed Centrifugation

Biomedicines ◽  
2020 ◽  
Vol 8 (8) ◽  
pp. 246 ◽  
Author(s):  
Anders Askeland ◽  
Anne Borup ◽  
Ole Østergaard ◽  
Jesper V. Olsen ◽  
Sigrid M. Lund ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Size Exclusion Chromatography ◽  
High Speed ◽  
Proteome Analysis ◽  
Extracellular Vesicle ◽  
Size Exclusion ◽  
Label Free ◽  
Protein Biomarkers ◽  
Transmission Electron ◽  
Exclusion Chromatography

Extracellular vesicles (EVs) are small membrane-enclosed particles released by cells under various conditions specific to cells’ biological states. Hence, mass-spectrometry (MS) based proteome analysis of EVs in plasma has gained much attention as a method to discover novel protein biomarkers. MS analysis of EVs in plasma is challenging and EV isolation is usually necessary. Therefore, we compared differences in abundance, subtypes, and contamination for EVs isolated by high-speed centrifugation, size exclusion chromatography (SEC), and peptide-affinity precipitation (PAP/ME kit) for subsequent MS-based proteome analysis. Successful EV isolation was evaluated by nanoparticle-tracking analysis, immunoblotting, and transmission electron microscopy, while EV abundance, EV subtypes, and contamination was evaluated by label-free tandem MS. High-speed centrifugation and SEC isolates showed high EV abundance at the expense of contamination by non-EV proteins and lipoproteins, respectively. These two methods also resulted in EVs of a similar type, however, with smaller EVs in SEC isolates. PAP isolates had a relatively low EV abundance and high contamination. We consider high-speed centrifugation and SEC suitable as EV isolation for MS biomarker studies, where the choice between the two should depend on the scientific questions and whether the focus is on larger or smaller EVs or a combination of both.

Size-exclusion chromatography in multidimensional separation schemes for proteome analysis

2003 ◽  
Vol 56 (1-3) ◽  
pp. 141-152 ◽  
Author(s):  
Paolo Lecchi ◽  
Anand R. Gupte ◽  
Ricardo E. Perez ◽  
Lyubov V. Stockert ◽  
Fred P. Abramson
Keyword(s):  
Size Exclusion Chromatography ◽  
Proteome Analysis ◽  
Size Exclusion ◽  
Exclusion Chromatography ◽  
Multidimensional Separation
Sign in / Sign up

Export Citation Format

Share Document