Introduction
This application note demonstrates the analysis of trypsin-digested bovine serum albumin (BSA) with on- column digestion using Tecan’s Narrow Bore Extraction (NBE) columns, followed by peptide clean-up by Solid Phase Extraction (SPE) utilizing Atlas® sorbent in the same column. Resulting recoveries were then compared to standard in-tube trypsin digestion followed by subsequent sample clean-up using the same Atlas sorbents in SPE columns.
NBE columns offer potential workflow advantages due to their unique anti-flow airgap between the reaction vessel and bottom chamber (which contains the SPE sorbent). This airgap prevents liquid from entering the sorbent chamber until positive pressure is applied. The empty upper chamber of the NBE column can be used independently of the lower chamber as a vessel for sample preparation. This upper chamber is widely used for assays utilizing enzymatic hydrolysis prior to SPE. The aim of these experiments is to demonstrate similar benefits when using NBE columns as the incubation vessel as well as the sample cleanup column for proteomics workflows, resulting in a simplified and fast sample processing procedure.
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