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High-throughput Platform (high-throughput + platform)
Selected AbstractsToxicological assessment of chemicals using Caenorhabditis elegans and optical oxygen respirometryENVIRONMENTAL TOXICOLOGY & CHEMISTRY, Issue 4 2009Katherine Schouest Abstract Oxygen consumption is indicative of an organism's metabolic state, whereby alterations in respiration rate can result from the presence of different stimuli. Here, we develop a novel approach based on quenched fluorescence oxygen sensing and respirometry method for toxicity screening assays using the nematode Caenorhabditis elegans. Previously, C. elegans was established as a useful model in soil and aquatic toxicology studies. For existing toxicology screening approaches with C. elegans, however, the endpoint is lethality. In addition, the assay time frame for the existing approaches is considerably longer than that for the approach described here. We present a sensitive, robust, high-throughput platform using standard laboratory equipment for toxicological studies by measuring respiration rate in C. elegans animals using a phosphorescent probe. [source] Proteomic identification of biomarkers related to Helicobacter pylori -associated gastroduodenal disease: Challenges and opportunitiesJOURNAL OF GASTROENTEROLOGY AND HEPATOLOGY, Issue 11 2008Ming-Shiang Wu Abstract Helicobacter pylori colonize the stomach of over half the world's population. While 80,90% H. pylori -infected individuals have clinically asymptomatic gastritis, 10,15% develop peptic ulcer, and 1,2% gastric malignancies. These variable clinical outcomes have led to an interest in prognostic indicators. The current disease paradigm suggests that host genetics and bacterial virulence both play important roles in modulating the final outcome of H. pylori infection. Elucidation of the interaction between host and bacterium is essential to clarify pathogenesis and to develop new strategies for prevention and treatment. Proteomic technology is a powerful tool for simultaneously monitoring proteins and protein variation on a large scale in biological samples. It has provided an unprecedented opportunity to survey a cell's translational landscape comprehensively, and the results may allow in-depth analyses of host and pathogen interactions. Using this high-throughput platform and taking advantage of complete sequences for both the H. pylori and the human genome in available databases, we have identified several crucial proteins that have pathogenic and prognostic potential. Among them, antibodies to AhpC and GroEs of H. pylori could be utilized for identification of patients who are at high risk of disease complications after H. pylori infection. Evolving proteomic technologies, together with appropriate clinical phenotyping and genotype information should enhance understanding of disease pathogenesis and lead to more precise prediction of variable disease outcomes. It will also facilitate development of biomarkers for diagnosis, treatment, and prevention of H. pylori infection. [source] Microfluidics-based electrospray ionization enhances the intrasource separation of lipid classes and extends identification of individual molecular species through multi-dimensional mass spectrometry: development of an automated high-throughput platform for shotgun lipidomicsRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 13 2008Xianlin Han Herein, we exploit the use of microfluidics and optimized Taylor cones for improved intrasource separation/selective ionization of lipid classes during electrospray ionization. Increased differential ionization of multiple phospholipid classes was achieved through microfluidics with chip-based ionization resulting in substantial enhancement of intrasource separation/selective ionization of phospholipid classes in comparison to the conventional ion source. For example, using myocardial lipid extracts, 3-fold improvements in intrasource separation/selective ionization of myocardial phospholipid classes were routinely realized in the negative-ion mode in the absence of LiOH or other basic modifiers in the infused sample solutions. Importantly, the relative ratios of ions corresponding to individual molecular species in each lipid class to a selected internal standard from myocardial extracts were nearly identical between the chip-based interface and the syringe-pump-driven capillary interface. Therefore, quantitation of individual lipid molecular species directly from biological extracts through comparisons with internal standards in each lipid class was readily accomplished with an accuracy and dynamic range nearly identical to those documented using the well-established direct syringe-pump-driven capillary interface. Collectively, the use of microfluidics and robotic sample handling substantially enhances intrasource separation of lipids in comparison to routine capillary interfaces and greatly facilitates the use of multi-dimensional mass spectrometry using shotgun lipidomics, thereby providing an automated and high-throughput platform for global analyses of cellular lipidomes. Copyright © 2008 John Wiley & Sons, Ltd. [source] High-throughput screening techniques for rapid PEG-based precipitation of IgG4 mAb from clarified cell culture supernatantBIOTECHNOLOGY PROGRESS, Issue 3 2010Carol Knevelman Abstract Locating optimal protein precipitation conditions for complex biological feed materials is problematic. This article describes the application of a series of high-throughput platforms for the rapid identification and selection of conditions for the precipitation of an IgG4 monoclonal antibody (mAb) from a complex feedstock using only microliter quantities of material. The approach uses 96-microwell filter plates combined with high-throughput analytical methods and a method for well volume determination for product quantification. The low material, time and resource requirements facilitated the use of a full factorial Design of Experiments (DoE) for the rapid investigation into how critical parameters impact the IgG4 precipitation. To aid the DoE, a set of preliminary range-finding studies were conducted first. Data collected through this approach describing Polyethylene Glycol (PEG) precipitation of the IgG4 as a function of mAb concentration, precipitant concentration, and pH are presented. Response surface diagrams were used to explore interactions between parameters and to inform selection of the most favorable conditions for maximum yield and purification. PEG concentrations required for maximum yield and purity were dependant on the IgG4 concentration; however, concentrations of 14 to 20% w/v, pH 6.5, gave optimal levels of yield and purity. Application of the high-throughput approach enabled 1,155 conditions to be examined with less than 1 g of material. The level of insights gained over such a short time frame is indicative of the power of microwell experimentation in allowing the rapid identification of appropriate processing conditions for key bioprocess operations. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2010 [source] | ||||||||||