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Petersmann A.,Universitatsmedizin Greifswald | Baum H.,Regionale Kliniken Holding RKH GmbH | Bietenbeck A.,TU Munich | Braun S.L.,Institute For Laboratoriumsmedizin | And 28 more authors.
LaboratoriumsMedizin | Year: 2015

The association Institut für Qualitätsmanagement in Medizinischen Laboratorien (INQUAM) promotes the principle of excellence and supports medical laboratories on their way to the first step of the quality management system of the European Foundation for Quality Management "Committed to Excellence" with an adapted matrix for self-evaluation of medical laboratories. © 2015 by De Gruyter.


Posch A.,Bio Rad Laboratories GmbH
Archives of Physiology and Biochemistry | Year: 2014

Sample preparation is one of the key technologies for successful two-dimensional electrophoresis (2DE). Due to the great diversity of protein sample types and sources, no single sample preparation method works with all proteins; for any sample the optimum procedure must be determined empirically. This review is meant to provide a broad overview of the most important principles in sample preparation in order to avoid a multitude of possible pitfalls. Sample preparation protocols from the expert in the field were screened and evaluated. On the basis of these protocols and my own comprehensive practical experience important guidelines are given in this review. The presented guidelines will facilitate straightforward protocol development for researchers new to gel-based proteomics. In addition the available choices are rationalized in order to successfully prepare a protein sample for 2DE separations. The strategies described here are not limited to 2DE and can also be applied to other protein separation techniques. © 2014 Informa UK Ltd.


Adams O.,Universitatsklinikum Dusseldorf | Cooper G.,W. Gregory Cooper LLC | Fraser C.,Scottish Bowel Screening Center | Hubmann M.,MZ Labor | And 4 more authors.
Clinical Chemistry and Laboratory Medicine | Year: 2012

In April of 2011, Bio-Rad Laboratories Quality System Division (Irvine, CA, USA) hosted its third annual convocation of experts on laboratory quality in the city of Salzburg, Austria. As in the past 2 years, over 60 experts from across Europe, Israel, USA and South Africa convened to discuss contemporary issues and topics of importance to the clinical laboratory. This year's conference had EN/ISO 15189 and accreditation as the common thread for most discussions, with topics ranging from how to meet requirements like uncertainty to knowledge gained from those already accredited. The participants were divided into five discussion working groups (WG) with assigned topics. The outcome of these discussions is the subject of this summary. © 2012 by Walter de Gruyter.


Posch A.,Bio Rad Laboratories GmbH | Franz T.,MPI for Biology of Ageing | Hartwig S.,Institute of Clinical Biochemistry | Knebel B.,Institute of Clinical Biochemistry | And 7 more authors.
Archives of Physiology and Biochemistry | Year: 2013

Two-dimensional gel electrophoresis (2-DE) is one of the most powerful methods for studying global protein profiles. However, due to the multiple manual steps involved in gel based processing it is challenging to achieve the necessary overall reproducibility for a reliable comparative analysis, especially between different laboratories. To improve the 2-DE technique for quantitative analyses we have set up a robust 2-DE workflow, called 2D-ToGo, which utilizes latest innovations concerning instrumentation, consumables and protocols. Quantitative data analyses indicate the high reproducibility between replicate gels processed at a single site (intra-laboratory variation: CV 20%). The data-sets of the inter-laboratory comparison revealed similar results displaying a variation of CV 23%. The technical improvements given by our 2-DE workflow have a positive impact on process robustness and most importantly, reproducibility. Accordingly, many of the well-known challenges for resolving and quantitating up to thousands of different protein components in a given biological sample are minimized. © 2013 Informa UK Ltd.


Taylor S.C.,Bio Rad Laboratories Inc. | Posch A.,Bio Rad Laboratories GmbH
BioMed Research International | Year: 2014

Western blotting is a technique that has been in practice for more than three decades that began as a means of detecting a protein target in a complex sample. Although there have been significant advances in both the imaging and reagent technologies to improve sensitivity, dynamic range of detection, and the applicability of multiplexed target detection, the basic technique has remained essentially unchanged. In the past, western blotting was used simply to detect a specific target protein in a complex mixture, but now journal editors and reviewers are requesting the quantitative interpretation of western blot data in terms of fold changes in protein expression between samples. The calculations are based on the differential densitometry of the associated chemiluminescent and/or fluorescent signals from the blots and this now requires a fundamental shift in the experimental methodology, acquisition, and interpretation of the data. We have recently published an updated approach to produce quantitative densitometric data from western blots (Taylor et al., 2013) and here we summarize the complete western blot workflow with a focus on sample preparation and data analysis for quantitative western blotting. © 2014 Sean C. Taylor and Anton Posch.


Haeckel R.,Bremer Zentrum For Laboratoriumsmedizin | Sonntag O.,Bio Rad Laboratories GmbH
LaboratoriumsMedizin | Year: 2012

The validation of analytical procedures in laboratory medicine has been discussed by many authors and several recommendations have been elaborated by working groups and standardization bodies. However, some problems have not sufficiently been solved, for example, permissible limits for the comparability between methods (permissible equivalence), what is the best statistical procedure for estimating the function of the fitting line in scatter plots, and permissible limits for the spread around the fitting line. The present article attempts to summarize the present state-of-the-art and to provide answers to some of the open problems. © 2012 by Walter de Gruyter Berlin Boston.


Moschallski M.,University of Tübingen | Hausmann M.,Bio Rad Laboratories GmbH | Posch A.,Bio Rad Laboratories GmbH | Paulus A.,Bio Rad Laboratories GmbH | And 10 more authors.
Electrophoresis | Year: 2010

We have developed a microfluidic system - microPrep - for subcellular fractionation of cell homogenates based on dielectrophoretic sorting. Separation of mitochondria isolated from a human lymphoblastoid cell line was monitored by fluorescence microscopy and further characterized by western blot analysis. Robust high throughput and continuous long-term operation for up to 60 h of the microPrep chip system with complex biological samples became feasible as a result of a comprehensive set of technical measures: (i) coating of the inner surfaces of the chip with BSA, (ii) application of mechanical actuators to induce periodic flow patterns, (iii) efficient cooling of the device to ensure integrity of organelle, (iv) a wide channel to provide for high fluidic throughput, and (v) integration of a serial arrangement of 10 dielectrophoretic deflector units to enable separation of samples with a high particle load without clogging. Hence, microPrep yields tens of micrograms of enriched and purified mitochondria within hours. Western blots of mitochondria fractions showed that contaminating endoplasmatic reticulum was reduced by a factor 6 when compared with samples prepared by state of the art centrifugation. © 2010 Wiley-VCH Verlag GmbH & Co. KGaA.


Sciacovelli L.,University of Padua | Sonntag O.,Bio Rad Laboratories GmbH | Padoan A.,University of Padua | Zambon C.F.,University of Padua | And 2 more authors.
Clinical Chemistry and Laboratory Medicine | Year: 2012

Background: Data on quality indicators (QIs) should be collected over time in order to identify and continuously monitor clinical laboratory performance and to improve patient safety by identifying and implementing effective interventions. The aim of the present study was to ascertain whether the utilization of a set of quality indicators over a 3-year period resulted in an improvement in the efficiency and effectiveness of an individual laboratory. Methods: Over a 3-year time interval (20092011), a series of 38 QIs covering all stages of the total testing process (21 in the pre-analytic, nine in the analytic and eight in the post-analytic phase) was monitored. Results: On the basis of their patterns, QIs have been grouped into the following categories: [1] seven QIs of the pre-analytical phase and three of the intra-analytical phase with a significant trend and a significant linearity demonstrating an improvement over time; [2] 10 QIs of the pre-analytical and two of the intra-analytical phase with a significant trend and a non-significant linearity demonstrating that changes were not constant; [3] two QIs of the pre-analytical and one of the intra-analytical phase with a non-significant trend and significant linearity showing neither improvement nor worsening; and [4] two QIs of the pre-analytical and three of the intra-analytical phase with a non-significant trend and non-significant linearity. Conclusions: Data on a set of QIs collected over a 3-year time-frame demonstrate that processes and indicators under the control of the clinical laboratory had improved much more than processes requiring close co-operation between the laboratory and care teams. © 2012 by Walter de Gruyter Berlin Boston.

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