Automated MALDI-TOF MS based high-throughput screening workflow for in vitro enzyme assays (IMAGE)

SLAS (Society for Laboratory Automation and Screening)

Automated MALDI-TOF MS based high-throughput screening workflow for in vitro enzyme assays

Caption

Liquid handling devices are used for (1) compound, (2) enzyme, (3) substrate (S), and (4) internal standard (iSt) in quench solution delivery which is interrupted by sensible incubation times (indicated above arrows) to allow compound enzyme alignment and product (P) formation over time. An isotopically labelled internal standard is spiked into the samples to allow quantification. (5) The sample is mixed with matrix and delivered onto the target plate which is acquired by MALDI-TOF MS. (6) Upon laser ionisation, analyte ions are separated in a flight tube according to their mass-to-charge (m/z) ratio before they hit a detector. (7) In the mass spectrum, active and inactive compounds can be distinguished based on the observed product formation.

Credit

Leonie Müller, et al.

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