Collagenase isoforms for pancreas digestion.
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 beta and another of 106 kDa) and one to class II (isoform delta) 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.
Bertuzzi
Federico
F
Unit of Cell Therapy for Type 1 Diabetes Mellitus, San Raffaele Scientific Institute, Milan, Italy.
Cainarca
Silvia
S
Marzorati
Simona
S
Bachi
Angela
A
Antonioli
Barbara
B
Nano
Rita
R
Verzaro
Roberto
R
Ricordi
Camillo
C
D017364:Collagenases / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D006801:Humans; D007527:Isoenzymes / Q000737:chemistry; D010179:Pancreas / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods; D013820:Thermolysin / Q000737:chemistry*
D007527:Isoenzymes; D017364:Collagenases; C106635:Liberase; D013820:Thermolysin