Highkin Maureen K, Yates Matthew P, Nemirovskiy Olga V, Lamarr William A, Munie Grace E, Rains John W, Masferrer Jaime L, Nagiec Marek M
Pfizer Global R&D, St. Louis, MO 63017, USA.
J Biomol Screen. 2011 Feb;16(2):272-7. doi: 10.1177/1087057110391656.
To facilitate discovery of compounds modulating sphingosine-1-phosphate (S1P) signaling, the authors used high-throughput mass spectrometry technology to measure S1P formation in human whole blood. Since blood contains endogenous sphingosine (SPH) and S1P, mass spectrometry was chosen to detect the conversion of an exogenously added 17-carbon-long variant of sphingosine, C17SPH, into C17S1P. The authors developed procedures to achieve homogeneous mixing of whole blood in 384-well plates and for a method requiring minimal manipulations to extract S1P from blood in 96- and 384-well plates prior to analyses using the RapidFire(®) mass spectrometry system.
为便于发现调节1-磷酸鞘氨醇（S1P）信号传导的化合物，作者使用高通量质谱技术来测定人全血中S1P的生成。由于血液中含有内源性鞘氨醇（SPH）和S1P，因此选择质谱法来检测外源性添加的17碳长鞘氨醇变体C17SPH向C17S1P的转化。作者开发了在384孔板中实现全血均匀混合的程序，以及一种在使用RapidFire®质谱系统进行分析之前，从96孔板和384孔板中的血液中提取S1P所需操作最少的方法。
