SpeedScreen is a novel, label-free, in-solution, affinity-based selection methodology for high-throughput screening (HTS) developed at Novartis Pharma. The SpeedScreen protocol comprises in-solution affinity selection, followed by size exclusion chromatography in combination with microbore-liquid-chromatography/electrospray-ionization mass spectrometry (micro-LC/ESI-MS). The authors describe the basic concept behind assay development, HTS, and data analysis with the SpeedScreen technology. Advantages and limitations of SpeedScreen compared to alternative screening technologies are discussed, and an example is given from a SpeedScreen campaign applying this innovative affinity selection concept in HTS.
1. Venter JC, Adams MD, Myers EW, Li PW, Mural RJ, Sutton GG, et al: The sequence of the human genome. Science2001;291:1304-1354.
2.
2. Muckenschnabel I, Falchetto R, Mayr LM, Filipuzzi I: SpeedScreen: label-free, liquid-chromatography-mass spectrometry-based high-throughput screening for the discovery of orphan protein ligands. Anal Biochem2004;324:241-249.
3.
3. Louris JN, Cooks RG, Syka JEP, Kelley PE, Stafford GC Jr, Todd JFJ: Instrumentation, applications, and energy deposition in quadrupole ion-trap tandem mass spectrometry. Anal Chem1987;59:1677-1685.
4.
4. Prade L, Engh RA, Girod A, Kinzel V, Huber R, Bossmeyer D: Staurosporine-induced conformational changes of c-AMP dependent protein kinase catalytic subunit explain inhibitory potential. Structure1997;5:1627-1637.
5.
5. Hershko A, Ciechanover A: The ubiquitin system. Annu Rev Biochem1998;67:425-479.
