Advanced Electrophoresis Solutions (Canada)

Animal-free (AF) SERVA Collagenase AF-1 and Neutral Protease (NP) AF GMP Grade have recently become available for human islet isolation. This report describes the initial experiences of 3 different islet transplant centers. Thirty-four human pancreases were digested using 1 vial of the 6 different lots of Collagenase AF-1 (2,000–2,583 PZ-U/vial) supplemented with 4 different lots of NP AF in a range of 50 to 160 DMC-U per pancreas. Isolation, culture, and quality assessment were performed using standard techniques as previously described. All data are presented as mean ± standard error of the mean (SEM). Variability of pancreas weight was associated with a wide range of collagenase and NP activities, ranging from 12.7 to 46.6 PZ-U/g (26.0 ± 1.5 PZ-U/g) and 0.4 to 3.0 DMC-U/g (1.5 ± 0.1 DMC-U/g), respectively. Postpurification islet yield was 296,494 ± 33,620 islet equivalents (IEQ) equivalent to 3,274 ± 450 IEQ/g with a purity of 55.9% ± 3.2%. Quality assessment performed after 2 to 4 d of culture demonstrated a viability of 88.1% ± 1.5% and a stimulation index of 3.7 ± 0.7. Eighteen of the 34 preparations were transplanted into type 1 diabetic patients equivalent to a transplantation rate of 52.9%. Six preparations, which were infused into patients as first transplant, could be analyzed and increased the fasting C-peptide level from 0.11 ± 0.08 pretransplant to 1.23 ± 0.24 and 2.27 ± 0.31 ng/mL 3 and 6 mo posttransplant (P < 0.05), respectively. Insulin requirements were simultaneously reduced at the same time from 39.2 ± 3.8 IU/d before transplantation to 10.8 ± 4.1 and 4.0 ± 2.3 IU/d, after 3 and 6 mo posttransplant (P < 0.05), respectively. This study demonstrates the efficiency of AF SERVA Collagenase AF-1 and NP AF for clinical islet isolation and transplantation. The new plant-based production process makes these products a safe new option for the islet field.

Today the omnipresence of MS-based proteomic approaches suggest, that 2-dimensional gel (2-DE) based proteomics is dead. In reality, this is far from the truth, and also after three decades high-resolution 2-DE, introduced by Patrick O’Farrell in 1975, still represents a key technology to investigate thousands of protein species, their isoforms and modifications simultaneously. Especially, in combination with multi fluorescence labelling and various MS-methodologies, it is very popular to relatively quantify and identify proteins in complex mixtures. But also after these 30 years of use, reproducibility and process stability are still under constant improvement to further enhance robustness and feasibility of the method. In order to offer an opportunity to discuss and apply the latest developments in 2-DE in a scientific context we picked out this technology as central theme for the International Protein Rainbow Workshop titled ‘‘2-Dimensional Gel Electrophoresis Reloaded’’, which was held at the Institute of Clinical Biochemistry and Pathobiochemistry at the German Diabetes-Center in Duesseldorf. This annual hands-on training course, comprising theoretical and practical aspects of the technology, was accompanied by a lecture day giving an overview of recent 2-DE applications in scientific research. In dependence on this the current issue ‘‘2-Dimensional Gel Electrophoresis Reloaded’’ should help to give an overview of recent developments and applications related to 2-DE. The collection of scientific reports covers a broad range of applications, starting with an inter-laboratory comparison introducing a simplified and robust 2-DE work-flow. In addition a novel fluorescent dye for improved protein detection is described and pre-analytical conditions for serum proteomics, representing one crucial point for biomarker discovery are addressed. Another study examines differences and similarities between protein patterns derived from mitochondria and peroxisomes or describe the characterization of sub-types in small cell lung cancer (SCLC).