Jagadeesan Kishore Kumar, Wierzbicka Celina, Laurell Thomas, Sellergren Börje, Shinde Sudhirkumar, Ekström Simon
Department of Biomedical Engineering, Lund University, Lund, Sweden.
Department of Biomedical Sciences, Faculty of Health and Society, Malmö University, Sweden.
J Chromatogr B Analyt Technol Biomed Life Sci. 2016 May 15;1021:213-220. doi: 10.1016/j.jchromb.2015.10.033. Epub 2015 Oct 30.
Technology that facilitates rapid investigation of solid phase extraction protocols using very small amounts of sorbent can save both time and money. The microfabricated ISET (Integrated Selective Enrichment Target) interfaced with MALDI mass spectrometry is able to provide an efficient, economic and generic optimization process for SPE sample preparation. The SPE is performed in a rapid and parallel fashion, with a processing time off only 2h per ISET with 96 samples. Each of the 96 wells on the ISET can hold 600nL of SPE sorbent. The ability to work with small amounts of sorbent and samples in the ISET platform provides a big advantage when developing affinity sorbents, such as molecularly imprinted polymers (MIPs). Here it is demonstrated that an amount of 25mg phosphoserine imprinted MIP (pS-MIP) sorbent can allow for analysis of more than 500 ISET nanovials using a multitude of different conditions. In the presented case, the multiplexed experiments allowed for early discovery of unspecific interactions and subsequent minimization of these, resulting in a protocol that provided improved enrichment of phosphopeptides.
能够使用极少量吸附剂快速研究固相萃取方案的技术可以节省时间和金钱。与基质辅助激光解吸电离质谱联用的微加工集成选择性富集靶标（ISET）能够为固相萃取样品制备提供高效、经济且通用的优化过程。固相萃取以快速并行的方式进行，每个ISET处理96个样品的处理时间仅为2小时。ISET上的96个孔中的每一个都可以容纳600纳升的固相萃取吸附剂。在ISET平台上使用少量吸附剂和样品的能力在开发亲和吸附剂（如分子印迹聚合物（MIP））时具有很大优势。在此证明，25毫克磷酸丝氨酸印迹MIP（pS-MIP）吸附剂的量可以在多种不同条件下对500多个ISET纳米小瓶进行分析。在本案例中，多重实验允许早期发现非特异性相互作用并随后将其最小化，从而产生一种能够改善磷酸肽富集的方案。
