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Characterizing Biotherapeutics with HPLC

Smart Solutions for large Molecules

24-11-2010

As demonstrated here, there are a broad range of HPLC applications used in development, production, validation and release of protein therapeutics for human use. Some of the applications mentioned above are mandatory when submitting a new biopharmaceutical to approval at the regulatory agencies. The introduction of the first so-called biosimilars in Europe further increased the demand for highly efficient analysis methods. The continuous development of new stationary phases for the common modes of biochromatography, such as size exclusion, ion exchange and hydrophilic interaction chromatography support biochemists in increasing lab productivity and establishing high-throughput HPLC analysis not only for small molecules but also for large biopolymers...

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High-Throughput Analysis Using the Novel LightCycler® 1536 Real-Time PCR System

07-09-2009

An important trend in biological and biochemical analysis over the past 15 years has been miniaturization and parallelization of analytical procedures. Large-scale high-throughput analysis of gene expression or genetic polymorphisms is an essential element of modern functional genomics, where individual samples are screened against many thousands of target genes. According to its superior sensitivity and accuracy, qPCR is a well-established gold standard for precise profiling of gene expression levels or genotyping of known SNPs...

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Transcriptor High Fidelity cDNA Synthesis Kit - Accuracy Meets RT-PCR

04-06-2008

Retroviral reverse transcriptases commonly used for cDNA synthesis exhibit a higher error rate than other DNA polymerases used in nucleic acid analysis techniques. This lack of accuracy leads to a significant number of base exchanges or frameshifts which are further propagated in a subsequent PCR reaction. With the introduction of a new high-accuracy reverse transcriptase, Roche Applied Science now offers a new tool to increase the fidelity of cDNA synthesis. The Transcriptor High Fidelity cDNA Synthesis Kit not only reduces the amount of errors in RT-PCRs; it also allows the reverse transcription of full-length cDNAs with high yield....

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Molecular Probes® NanoOrange® Assay Performed on BMG LABTECH FLUOstar OPTIMA Microplate Reader

08-05-2008

The field of proteomics has expanded dramatically in recent years with research on a whole host of organisms. Techniques such as two dimensional difference gel electrophoresis (2D DIGE) require the accurate quantitation of protein, as up to three labelled samples can be loaded on a single gel for comparison. Assay methods for determining protein quantitation include absorbance at 280 nm(1), the Bradford assay(2), Lowry assay(3), BCA method(4) and more sensitive assays such as Fluoroprofile® (Sigma-Aldrich) and NanoOrange® (Molecular Probes®)5()....

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Real-Time PCR Quantification of Plant miRNAs Using Universal ProbeLibrary Technology

29-08-2007

MicroRNAs (miRNAs) are naturally occurring, highly conserved families of transcripts (18-25 nt in length) that are processed from larger hairpin precursors. Both plant and animal genomes encode miRNAs, and to date, there are over 4000 mature miRNA transcripts annotated. Although miRNAs represent a relatively abundant class of transcripts, their expression levels vary greatly among species and tissues. Less abundant miRNAs routinely fail detection using technologies such as cloning, northern hybridization and microarray analysis. Here, we present a real-time quantification method for accurate, sensitive, and economical detection of plant miRNAs based upon the stem-loop RT primer approach using an miRNA-specific primer coupled with a stem-loop specific Universal ProbeLibrary (UPL) probe. This approach represents a highly specific, sensitive, and easy-to-apply system to detect miRNAs....

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Quantitative Detection of Legionella pneumophila in Water Samples: Assay Transfer to the LightCycler® 480 Real-Time PCR System

22-08-2007

An assay to detect and quantify Legionella spp. in water samples was successfully transferred from a capillary-based LightCycler® Instrument to the novel multiwell plate-based LightCycler® 480 System. A previously described protocol was easily adapted by modifying the fluorescent label of the detection probe to match the filter set of the LightCycler® 480 Instrument. While reproducibility and analytical sensitivity of the assay were found to be comparable on both systems, the novel plate-based instrument allowed for higher sample throughput and reduced assay time per sample....

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The Determination of Hazardous Substances According to RoHS and WEEE Directives

28-08-2005

1st July 2006 is an important deadline for the electro- and electronics industries. This date bans hazardous substances like lead, cadmium, mercury, hexavalent chromium and polybrominated biphenyls and diphenyl ethers from electro- and electronic equipment. This restriction, even though in discussion since 1991, has become more and more important from year to year because of tons of electronic waste releasing toxic substances which are accumulated in the environment, the food chain and all human beings. In order to enforce the substances ban and the limitation or substitution of hazardous components, elemental analysis is obviously the most important control measure for monitoring limiting values. This requires precise analytical systems such as X-ray fluorescence, ICP- and atomic absorption spectrometers. These instruments are able to detect trace concentrations of hazardous compounds - for example cadmium, using an atomic absorption spectrometer in the flame atomization mode up to 0.1 mg/L, or using the di...

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