The separation of pancreatic procarboxypeptidases by high-performance liquid chromatography and chromatofocusing

Francisco J. Burgos, Roger Pascual, Josep Vendrell, Claudi M. Cuchillo, Francesc X. Avilés

Research output: Contribution to journalArticleResearchpeer-review

5 Citations (Scopus)

Abstract

Different experimental conditions and chromatographic supports have been selected for the most efficient and rapid purification of procarboxypeptidases from porcine and human pancreas by different high-performance liquid chromatography (HPLC) variants (anion exchange, reversed phase and gel filtration). Anion-exchange chromatography was found to be the most capable and permitted the isolation, in a single step, of three different porcine procarboxypeptidases (2A + 1B forms) and five different human procarboxypeptidases (2B + 3A forms) in a native and pure state from whole pancreas extracts. Other pancreatic proproteases are also cleanly isolated in the same step. Reversed-phase chromatography under mild conditions separated porcine or human procarboxypeptidases A from other pancreatic proteins in a very short time but was unable further to subfractionate the same proteins. The sequential use of gel filtration (or anion-exchange) and reversed-phase HPLC chromatography permitted, in a simple way, the isolation and dissociation of the strongly bound components of the binary complexes between procarboxypeptidases A and proproteinase E in either porcine or human pancreas extracts. Chromatofocusing on a fast protein liquid chromatographic support was also found to be a very efficient technique, showing a slightly lower capability to separate procarboxypeptidases than anion-exchange HPLC though in a much shorter time and in larger quantities. © 1989.
Original languageEnglish
Pages (from-to)233-243
JournalJournal of Chromatography A
Volume481
Issue numberC
DOIs
Publication statusPublished - 1 Jan 1989

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