Collagenase Isoforms for Pancreas Digestion

被引：19

作者：

Bertuzzi, Federico ^{[6
]}

Cainarca, Silvia ^{[3
]}

Marzorati, Simona ^{[2
]}

Bachi, Angela ^{[4
]}

Antonioli, Barbara ^{[2
]}

Nano, Rita ^{[2
]}

Verzaro, Roberto ^{[5
]}

Ricordi, Camillo ^{[1
]}

机构：

[1] Univ Miami, Diabet Res Inst, Miami, FL 33136 USA

[2] Ist Sci San Raffaele, Unit Cell Therapy Type Diabet Mellitus 1, I-20132 Milan, Italy

[3] Axxam SpA, Milan, Italy

[4] Ist Sci San Raffaele, DIBIT, I-20132 Milan, Italy

[5] ISMETT, Dept Surg, Palermo, Italy

[6] Osped Niguarda Ca Granda, Dept Diabet, Milan, Italy

来源：

CELL TRANSPLANTATION | 2009年 / 18卷 / 02期

关键词：

Islet isolation; Purification; Collagenase; Proteases; HUMAN ISLET ISOLATION; CLOSTRIDIUM-HISTOLYTICUM; INDIVIDUAL COLLAGENASES; CLASS-II; TRANSPLANTATION; ENZYMES; SUCCESS; BLENDS; RATIO;

D O I：

10.3727/096368909788341270

中图分类号：

Q813 [细胞工程];

学科分类号：

摘要：

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.

引用

页码：203 / 206

页数：4

共 16 条

[11] Islet isolation for allotransplantation: variables associated with successful islet yield and graft function [J].

Nano, R ;

Clissi, B ;

Melzi, R ;

Calori, G ;

Maffi, P ;

Antonioli, B ;

Marzorati, S ;

Aldrighetti, L ;

Freschi, M ;

Grochowiecki, T ;

Socci, C ;

Secchi, A ;

Di Carlo, V ;

Bonifacio, E ;

Bertuzzi, F .

DIABETOLOGIA, 2005, 48 (05) :906-912

[12] Requirements for success in clinical islet transplantation [J].

Ricordi, C ;

Inverardi, L ;

Kenyon, NS ;

Goss, J ;

Bertuzzi, F ;

Alejandro, R .

TRANSPLANTATION, 2005, 79 (10) :1298-1300

[13] Mass spectrometric sequencing of proteins from silver stained polyacrylamide gels [J].

Shevchenko, A ;

Wilm, M ;

Vorm, O ;

Mann, M .

ANALYTICAL CHEMISTRY, 1996, 68 (05) :850-858

[14] Deterioration and variability of highly purified collagenase blends used in clinical islet isolation [J].

Yamamoto, Toshiyuki ;

Ricordi, Camillo ;

Messinger, Shari ;

Sakuma, Yasunaru ;

Miki, Atsushi ;

Rodriguez, Rayner ;

Haertter, Ross ;

Khan, Aisha ;

Alejandro, Rodolfo ;

Ichii, Hirohito .

TRANSPLANTATION, 2007, 84 (08) :997-1002

[15]

YOSHIDA ETUSUKO, 1965, BIOCHIM BIOPHYSACTA, V105, P562

[16] Profound: An expert system for protein identification using mass spectrometric peptide mapping information [J].

Zhang, WZ ;

Chait, BT .

ANALYTICAL CHEMISTRY, 2000, 72 (11) :2482-2489

← 1 2 →