Kinetic Capillary Electrophoresis with Mass-Spectrometry Detection (KCE-MS) Facilitates Label-Free Solution-Based Kinetic Analysis of Protein-Small Molecule Binding

ChemBioChem ◽  
2011 ◽  
Vol 12 (17) ◽  
pp. 2551-2554 ◽  
Author(s):  
Jiayin Bao ◽  
Svetlana M. Krylova ◽  
Derek J. Wilson ◽  
Oren Reinstein ◽  
Philip E. Johnson ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Capillary Electrophoresis ◽  
Kinetic Analysis ◽  
Small Molecule ◽  
Free Solution ◽  
Label Free ◽  
Mass Spectrometry Detection ◽  
Kinetic Capillary Electrophoresis

Pre-equilibration kinetic size-exclusion chromatography with mass spectrometry detection (peKSEC-MS) for label-free solution-based kinetic analysis of protein–small molecule interactions

The Analyst ◽  
2015 ◽  
Vol 140 (4) ◽  
pp. 990-994 ◽  
Author(s):  
Jiayin Bao ◽  
Svetlana M. Krylova ◽  
Leonid T. Cherney ◽  
J. C. Yves Le Blanc ◽  
Patrick Pribil ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Kinetic Analysis ◽  
Small Molecule ◽  
Size Exclusion ◽  
Free Solution ◽  
Label Free ◽  
Mass Spectrometry Detection ◽  
Reversible Binding ◽  
Exclusion Chromatography ◽  
Molecule Interactions

Label-free solution-based kinetic analysis of reversible binding between protein and small molecule.

scholarly journals Affinity Capture Enrichment versus Affinity Depletion: A Comparison of Strategies for Increasing Coverage of Low-Abundant Human Plasma Proteins

2020 ◽  
Vol 21 (16) ◽  
pp. 5903
Author(s):  
Nicolai Bjødstrup Palstrøm ◽  
Lars Melholt Rasmussen ◽  
Hans Christian Beck
Keyword(s):  
Mass Spectrometry ◽  
Small Molecule ◽  
Plasma Proteins ◽  
Biomarker Discovery ◽  
Plasma Samples ◽  
Label Free ◽  
Affinity Capture ◽  
Liquid Chromatography Tandem Mass ◽  
Removal System ◽  
Equalization Method

In the present study, we evaluated four small molecule affinity-based probes based on agarose-immobilized benzamidine (ABA), O-Phospho-L-Tyrosine (pTYR), 8-Amino-hexyl-cAMP (cAMP), or 8-Amino-hexyl-ATP (ATP) for their ability to remove high-abundant proteins such as serum albumin from plasma samples thereby enabling the detection of medium-to-low abundant proteins in plasma samples by mass spectrometry-based proteomics. We compared their performance with the most commonly used immunodepletion method, the Multi Affinity Removal System Human 14 (MARS14) targeting the top 14 most abundant plasma proteins and also the ProteoMiner protein equalization method by label-free quantitative liquid chromatography tandem mass spectrometry (LC-MSMS) analysis. The affinity-based probes demonstrated a high reproducibility for low-abundant plasma proteins, down to picomol per mL levels, compared to the Multi Affinity Removal System (MARS) 14 and the Proteominer methods, and also demonstrated superior removal of the majority of the high-abundant plasma proteins. The ABA-based affinity probe and the Proteominer protein equalization method performed better compared to all other methods in terms of the number of analyzed proteins. All the tested methods were highly reproducible for both high-abundant plasma proteins and low-abundant proteins as measured by correlation analyses of six replicate experiments. In conclusion, our results demonstrated that small-molecule based affinity-based probes are excellent alternatives to the commonly used immune-depletion methods for proteomic biomarker discovery studies in plasma. Data are available via ProteomeXchange with identifier PXD020727.

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