MS and HPLC Improvements

Besides using the most recently developed ESI mass spectrometers with sensitivity and resolution improvements of 10x and 3x, respectively, principally with time-of-flight mass spectrometers, the most significant improvements in GPC spin column/HPLC ESI-MS screening will come from the application of ultraperformance liquid chromatography (UPLC) where the combination of a very high pressure liquid chromatograph together with LC columns containing particles of 1.7 mm has been demonstrated to produce chromatograms of remarkably improved resolution (peak widths of 5 s) with retention times also reduced by a factor of up to 10 times and with high sensitivity, generally 3-10 x more than with conventional LC columns (Waters Acquity UPLC System) [37]. With these improvements in both LC and MS instrumentation, factors in sensitivity of 30100 x could be achieved vs presently used instrumentation, permitting more reliable detection of weaker signals in screening weak binders and enabling the use of smaller amounts of precious protein in a screening program, all achievable in considerably reduced cycle times.

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