Cell Transplantation (Feb 2009)

Collagenase Isoforms for Pancreas Digestion

  • Federico Bertuzzi,
  • Silvia Cainarca,
  • Simona Marzorati,
  • Angela Bachi,
  • Barbara Antonioli,
  • Rita Nano,
  • Roberto Verzaro,
  • Camillo Ricordi

DOI
https://doi.org/10.3727/096368909788341270
Journal volume & issue
Vol. 18

Abstract

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The available information concerning the characteristics and composition of collagenase batches, which are effective in the digestion of human pancreas for islet transplants, is scarce and incomplete. A large inter-and intrabatched variability in activity and efficiency of blend enzymes available for isolation has been observed. The aim of this study was to characterize enzyme blend components. Liberase batches were characterized by SDS-PAGE analyses, microelectrophoresis, and then by MALDI-TOF MS analysis. Three main bands were detected by SDS-PAGE analysis and submitted to MALDI-TOF MS analysis. Two bands were found to correspond to class I (isoform β and another of 106 kDa) and one to class II (isoform δ) collagenase. These results represent an important step towards a complete characterization of enzymes, with the final aim of identifying key components for a standardized product.