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Updated: Jun 26, 2026
A Spin-Tip Enrichment Strategy for Simultaneous Analysis of N-Glycopeptides and Phosphopeptides from Human Pancreatic Tissues
Published on: May 4, 2022
Extraction of trypsin from bovine pancreas by applying polyethyleneglycol/sodium citrate aqueous two-phase systems
Gisela Tubio1, Guillermo A Picó, Bibiana B Nerli
1Chemical Physics Department, CONICET, FonCyT, Faculty of Biochemical and Pharmaceutical Sciences, National University of Rosario, Rosario, Argentina.
Abstract:
The goal of this work was to determine the optimal conditions for separating trypsin (TRP) from alpha-chymotrypsin (ChTRP) and to apply them for trypsin purification from bovine pancreas by liquid-liquid extraction with polyethyleneglycol/sodium citrate (PEG/NaCit) aqueous two-phase systems. Partitioning behaviours of TRP and ChTRP are demonstrated to be very sensitive to variables such as PEG molecular weight, pH and tie line length. Aqueous two-phase systems (ATPSs) formed by PEG of MW 3350 and NaCit pH 5.20 showed the best separation capability. The addition of NaCl up to a final concentration of 7% (w/w) and the decrease of top/bottom volume ratio to 0.1 led to the recovery of 60% of pancreatic TRP in a concentrated form in the top phase with a 3-fold purification. Biomass presence up to 25% (w/w) of the total system mass did not affect significantly yield and purification parameters.
