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Purification Technique (purification + technique)
Selected AbstractsTesting of the influenza virus purification by CIEFELECTROPHORESIS, Issue 2 2010Marie Horká Abstract In virological practice, the pre-concentration, purification and subsequent determination of the purity and concentration of the viruses from the cultural medium and/or from the real sample are required. The conventional techniques used today are equipment demanding, time-consuming and laborious. In this study, the CIEF of influenza viruses with UV detection has been developed and subsequently used to test the purification of the virus from the biological samples. The equine and swine influenza viruses present in infected allantoic fluid of specific pathogen free embryonated chicken eggs were precipitated by using PEG 6000 and sodium chloride. The precipitated viruses were centrifuged at 14,000×g, and the impurities of different densities were removed by using the sucrose gradients. The efficiency of the virus purification technique was examined by the CIEF and compared to the results of real-time PCR. The pIs of both influenza viruses were determined. Simultaneously, the CIEF was found to be a suitable method for the rapid testing of the efficiency of the virus purification. [source] Single-step purification of the recombinant green fluorescent protein from intact Escherichia coli cells using preparative PAGEELECTROPHORESIS, Issue 17 2009Few Ne Chew Abstract Mechanical and non-mechanical breakages of bacterial cells are usually the preliminary steps in intracellular protein purification. In this study, the recombinant green fluorescent protein (GFP) was purified from intact Escherichia coli cells using preparative PAGE. In this purification process, cells disruption step is not needed. The cellular content of E. coli was drifted out electrically from cells and the negatively charged GFP was further electroeluted from polyacrylamide gel column. SEM investigation of the electrophoresed cells revealed substantial structural damage at the cellular level. This integrated purification technique has successfully recovered the intracellular GFP with a yield of 82% and purity of 95%. [source] Rapid production of a plasmid DNA encoding a malaria vaccine candidate via amino-functionalized poly(GMA- co -EDMA) monolithAICHE JOURNAL, Issue 11 2008Michael K. Danquah Abstract Malaria is a global health problem; an effective vaccine is urgently needed. Due to the relative poverty and lack of infrastructure in malaria endemic areas, DNA-based vaccines that are stable at ambient temperatures and easy to formulate have great potential. While attention has been focused mainly on antigen selection, vector design and efficacy assessment, the development of a rapid and commercially viable process to manufacture DNA is generally overlooked. We report here a continuous purification technique employing an optimized stationary adsorbent to allow high-vaccine recovery, low-processing time, and, hence, high-productivity. A 40.0 mL monolithic stationary phase was synthesized and functionalized with amino groups from 2-Chloro-N,N-diethylethylamine hydrochloride for anion-exchange isolation of a plasmid DNA (pDNA) that encodes a malaria vaccine candidate, VR1020-PyMSP4/5. Physical characterization of the monolithic polymer showed a macroporous material with a modal pore diameter of 750 nm. The final vaccine product isolated after 3 min elution was homogeneous supercoiled plasmid with gDNA, RNA and protein levels in keeping with clinical regulatory standards. Toxicological studies of the pVR1020-PyMSP4/5 showed a minimum endotoxin level of 0.28 EU/mg pDNA. This cost-effective technique is cGMP compatible and highly scalable for the production of DNA-based vaccines in commercial quantities, when such vaccines prove to be effective against malaria. © 2008 American Institute of Chemical Engineers AIChE J, 2008 [source] Technical note: Removal of metal ion inhibition encountered during DNA extraction and amplification of copper-preserved archaeological bone using size exclusion chromatographyAMERICAN JOURNAL OF PHYSICAL ANTHROPOLOGY, Issue 2 2009Carney D. Matheson Abstract A novel technique for the removal of metal ions inhibiting DNA extraction and PCR of archaeological bone extracts is presented using size exclusion chromatography. Two case studies, involving copper inhibition, demonstrate the effective removal of metal ion inhibition. Light microscopy, SEM, elemental analysis, and genetic analysis were used to demonstrate the effective removal of metal ions from samples that previously exhibited molecular inhibition. This research identifies that copper can cause inhibition of DNA polymerase during DNA amplification. The use of size exclusion chromatography as an additional purification step before DNA amplification from degraded bone samples successfully removes metal ions and other inhibitors, for the analysis of archaeological bone. The biochemistry of inhibition is explored through chemical and enzymatic extraction methodology on archaeological material. We demonstrate a simple purification technique that provides a high yield of purified DNA (>95%) that can be used to address most types of inhibition commonly associated with the analysis of degraded archaeological and forensic samples. We present a new opportunity for the molecular analysis of archaeological samples preserved in the presence of metal ions, such as copper, which have previously yielded no DNA results. Am J Phys Anthropol, 2009. © 2009 Wiley-Liss, Inc. [source] Continuous Venovenous Renal Replacement Therapy With a Pulsatile Tubular Blood Pump: Analysis of Efficacy ParametersARTIFICIAL ORGANS, Issue 1 2006Jesús López-Herce Abstract:, A three-valve pulsatile tubular pump was used in 10 pigs weighing 9.9 ± 0.3 kg, connected to a neonatal hemofiltration (HF) circuit to analyze the parameters to determine the ultrafiltration effectiveness for venovenous continuous renal replacement therapy. Forty 30-min periods were studied to analyze the influence of the catheter diameter, purification technique (HF or hemodiafiltration), pump pulsation frequency, percentage time in diastole, percentage time in diastole in which the postpump valve was closed, and percentage time in systole in which the postfilter valve was closed, on ultrafiltrate volume and blood flow through the filter. A higher ultrafiltrate flow was achieved with a frequency of 60 bpm than with 30 bpm, a diastolic time of 65% of the total cycle versus 50%, and a blood flow of greater than 20 mL/min versus less than 20 mL/min. The ultrafiltrate flow increases in relation to the blood flow, the frequency of the pump, and the diastolic time. [source] | ||||||||||