Jagadeesan KK, Wierzbicka C, Laurell T, Sellergren B, Shinde S, Ekström S
Journal of Chromatography B 1021 (-) 213-220 [2016-05-15; online 2015-10-30]
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.
Structural Proteomics [Service]