Bioaffinity Screening with a Rapid and Sample-Efficient Autosampler for Native Electrospray Ionization Mass Spectrometry
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. Mehr anzeigen
Persistenter Link
https://doi.org/10.3929/ethz-b-000507526Publikationsstatus
publishedExterne Links
Zeitschrift / Serie
Analytical ChemistryBand
Seiten / Artikelnummer
Verlag
American Chemical SocietyOrganisationseinheit
03430 - Zenobi, Renato / Zenobi, Renato
03852 - Schneider, Gisbert / Schneider, Gisbert
Förderung
178765 - Soft ionization mass spectrometry for studying noncovalent interactions (SNF)
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Is supplemented by: https://doi.org/10.3929/ethz-b-000494717