Bioaffinity Screening with a Rapid and Sample-Efficient Autosampler for Native Electrospray Ionization Mass Spectrometry
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Open access
Date
2021-10-05Type
- Journal Article
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. Show more
Permanent link
https://doi.org/10.3929/ethz-b-000507526Publication status
publishedExternal links
Journal / series
Analytical ChemistryVolume
Pages / Article No.
Publisher
American Chemical SocietyOrganisational unit
03430 - Zenobi, Renato / Zenobi, Renato
03852 - Schneider, Gisbert / Schneider, Gisbert
Funding
178765 - Soft ionization mass spectrometry for studying noncovalent interactions (SNF)
Related publications and datasets
Is supplemented by: https://doi.org/10.3929/ethz-b-000494717
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