Immobilized β-secretase on magnetic microparticles, a promising alternative for kinetic and inhibition assays
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| Year of publication | 2016 |
| Type | Conference abstract |
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| Description | Immobilization of enzymes on solid supports is a well-established strategy providing numerous benefits. In our work we used magnetic microparticles to immobilize clinically important protease, the ß secretase (BACE), which plays key role in the Alzheimer’s disease (AD) pathogenesis. Since curing AD is considered one of the biggest challenge modern society must face, great afford is being put into search for suitable inhibitors both in academia and pharmaceutical industry, concurrently rising demand for high-throughput, sensitive and cost effective methods. Nowadays Förster resonance energy transfer based assays are used almost exclusively to study BACE kinetics and inhibition. These assays offer very high throughput and sensitivity, but suffer from numerous inherited drawbacks. Capillary electrophoresis coupled to mass spectrometry (CE-MS) is an excellent alternative combing great features of both mass spectrometry, enabling use of unlabelled substrates with high sensitivity, and capillary electrophoresis, which requires miniscule sample volumes and short analysis time. Therefore CE-MS was used in our study to determine BACE activity. |
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