BACKGROUND: Efficient islet isolation represents a necessary requirement for successful islet transplantation as a treatment for type 1 diabetes. The choice of collagenase for pancreas digestion is critical for the isolation outcome, and Liberase is the most widely used enzyme, although large intra-batched variability in activity and efficiency has been observed. METHODS: The aim of this study was to characterize Liberase components and their relative role in pancreas digestion. Liberase batches were characterized by microelectrophoresis. RESULTS: By means of microelectrophoresis, we identified three main proteins each with different prevalences between batches. Two proteins were found to correspond to class I (CI) and one to class II (CII) collagenase. In a series of 163 islet isolations, we observed that the CII correlated with islet yield (P<0.001) and digestion time (P<0.001); additionally, CI directly correlated with purity (P=0.028). Finally, when CII and one of the CI isoforms were >50 percentile, 15 of 36 preparations were transplanted, with 27 of 127 transplanted in the other cases (P=0.013). CONCLUSION: These results represent an important step toward the characterization of enzymes, with the final aim of identifying key components for a standardized product

Characterization of collagenase blend enzymes for human islet transplantation / B. Antonioli, I. Fermo, S. Cainarca, S. Marzorati, R. Nano, M. Baldissera, A. Bachi, R. Paroni, C. Ricordi, F. Bertuzzi. - In: TRANSPLANTATION. - ISSN 0041-1337. - 84:12(2007 Dec 27), pp. 1568-1575. [10.1097/01.tp.0000295719.88525.60]

Characterization of collagenase blend enzymes for human islet transplantation

S. Marzorati;R. Paroni;
2007

Abstract

BACKGROUND: Efficient islet isolation represents a necessary requirement for successful islet transplantation as a treatment for type 1 diabetes. The choice of collagenase for pancreas digestion is critical for the isolation outcome, and Liberase is the most widely used enzyme, although large intra-batched variability in activity and efficiency has been observed. METHODS: The aim of this study was to characterize Liberase components and their relative role in pancreas digestion. Liberase batches were characterized by microelectrophoresis. RESULTS: By means of microelectrophoresis, we identified three main proteins each with different prevalences between batches. Two proteins were found to correspond to class I (CI) and one to class II (CII) collagenase. In a series of 163 islet isolations, we observed that the CII correlated with islet yield (P<0.001) and digestion time (P<0.001); additionally, CI directly correlated with purity (P=0.028). Finally, when CII and one of the CI isoforms were >50 percentile, 15 of 36 preparations were transplanted, with 27 of 127 transplanted in the other cases (P=0.013). CONCLUSION: These results represent an important step toward the characterization of enzymes, with the final aim of identifying key components for a standardized product
Islet isolation ; Purification ; Collagenase proteases ; Islet transplantation
Settore BIO/10 - Biochimica
27-dic-2007
Article (author)
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/35042
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