Bioaffinity Screening with a Rapid and Sample-Efficient Autosampler for Native Electrospray Ionization Mass Spectrometry
- 21 September 2021
- journal article
- research article
- Published by American Chemical Society (ACS) in Analytical Chemistry
- Vol. 93 (39), 13342-13350
- https://doi.org/10.1021/acs.analchem.1c03130
Abstract
Fast and efficient handling of ligands and biological targets are required in bioaffinity screening based on native electrospray ionization mass spectrometry (ESI-MS). We use a prototype microfluidic autosampler, called the “gap sampler”, to sequentially mix and electrospray individual small molecule ligands together with a target protein and compare the screening results with data from thermal shift assay and surface plasmon resonance. In a first round, all three techniques were used for a screening of 110 ligands against bovine carbonic anhydrase II, which resulted in five mutual hits and some false positives with ESI-MS presumably due to the high ligand concentration or interferences from dimethyl sulfoxide. In a second round, 33 compounds were screened in lower concentrations and in a less complex matrix, resulting in only true positives with ESI-MS. Within a cycle time of 30 s, dissociation constants were determined within an order of magnitude accuracy consuming only 5 pmol of ligand and less than 15 pmol of protein per screened compound. In a third round, dissociation constants of five compounds were accurately determined in a titration experiment. Thus, the gap sampler can rapidly and efficiently be used for high-throughput screening.Keywords
Funding Information
- Schweizerischer Nationalfonds zur F??rderung der Wissenschaftlichen Forschung (200020_178765)
This publication has 41 references indexed in Scilit:
- A cross-platform toolkit for mass spectrometry and proteomicsNature Biotechnology, 2012
- Biophysical techniques for ligand screening and drug designCurrent Opinion in Pharmacology, 2009
- Label‐free determination of protein–ligand binding constants using mass spectrometry and validation using surface plasmon resonance and isothermal titration calorimetryJournal of Molecular Recognition, 2009
- Applications of ESI-MS in drug discovery: interrogation of noncovalent complexesNature Reviews Drug Discovery, 2006
- DMSO-Related Effects in Protein CharacterizationSLAS Discovery, 2006
- High Throughput Screening via Mass Spectrometry: A Case Study Using AcetylcholinesteraseASSAY and Drug Development Technologies, 2004
- Automated Nano-Electrospray Mass Spectrometry for Protein-Ligand Screening by Noncovalent Interaction Applied to Human H-FABP and A-FABPSLAS Discovery, 2003
- Quantitative determination of noncovalent binding interactions using soft ionization mass spectrometryInternational Journal of Mass Spectrometry, 2002
- A Simple Statistical Parameter for Use in Evaluation and Validation of High Throughput Screening AssaysSLAS Discovery, 1999
- Tandem Mass SpectrometryScience, 1981