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71.
Oscar Salas-Solano Kunnel Babu SungAe Suhr Park Xinfeng Zhang Li Zhang Zoran Sosic Boris Boumajny Ming Zeng Kuang-Chuan Cheng Angelia Reed-Bogan Stacey Cummins-Bitz David A. Michels Monica Parker Paulina Bonasia Mingfang Hong Steven Cook Margaret Ruesch David Lamb Dora Bolyan Steffen Kiessig Darren Allender Brian Nunnally 《Chromatographia》2011,73(11-12):1137-1144
Interlaboratory comparisons are essential to bringing emerging technologies into biopharmaceutical industry practice and regulatory acceptance. As a result, an international team including 12 laboratories from 10 independent biopharmaceutical companies in the United States and Switzerland was formed to evaluate the precision and robustness of capillary isoelectric focusing (CIEF) to assess the charge heterogeneity of monoclonal antibodies. The different laboratories determined the apparent pI and the relative distribution of the charge isoforms of a representative monoclonal antibody (rMAb) sample using the same CIEF method. Statistical evaluation of the data was performed to determine within and between-laboratory consistencies and outlying information. The apparent pI data generated for each charge variant peak showed very good precision between laboratories with percentage of RSD values of ??0.5%. Similarly, the percentage of RSD for the rMAb charge variants percent peak area values are ??4.4% across different laboratories with different analysts using different lots of ampholytes and multiple instruments. Taken together, these results validate the appropriate use of CIEF in the biopharmaceutical industry in support of regulatory submissions. 相似文献
72.
An efficient, easy to perform, one-pot reaction cascade for the synthesis of 2,5-disubstituted pyrroles from p-toluenesulfonyl protected imines and 4-nitrobut-1-yne under a combination of base and gold(III) catalysis is reported. 相似文献
73.
Three new [2]rotaxanes with aromatic nitrogen donors appended to the crown ether wheel have been synthesized and used as ligands to coordinate Cd(II) ions. One of these yields a new type of 2-periodic, metal organic rotaxane framework in which the wheel rather than the axle is used to link the metal nodes. 相似文献
74.
75.
Hierarchical ZnO/Cu "corn-like" materials with high photodegradation and antibacterial capability under visible light 总被引:1,自引:0,他引:1
A novel hierarchical ZnO/Cu "corn-like" material was successfully synthesized via facile low temperature hydrothermal and photo-deposition methods. This novel hierarchical material was well characterized by FESEM, XRD, EDS, UV-Vis. This novel material displayed high photodegradation and antibacterial capability under the irradiation of visible light. This is owing to its unique properties, including maximized light utilization rate via red-shifting the light absorption spectrum, enhanced specific surface area for mass transfer and reaction sites, and retarded recombination of photogenerated electrons and holes. Moreover, this novel hierarchical ZnO/Cu "corn-like" material demonstrated easy recovery from treated water for reuse. All these excellent properties indicate that this novel hierarchical ZnO/Cu "corn-like" material has a bright future in water purification applications. 相似文献
76.
Zhong Q Bhattacharya S Kotsopoulos S Olson J Taly V Griffiths AD Link DR Larson JW 《Lab on a chip》2011,11(13):2167-2174
Quantitative polymerase chain reactions (qPCR) based on real-time PCR constitute a powerful and sensitive method for the analysis of nucleic acids. However, in qPCR, the ability to multiplex targets using differently colored fluorescent probes is typically limited to 4-fold by the spectral overlap of the fluorophores. Furthermore, multiplexing qPCR assays requires expensive instrumentation and most often lengthy assay development cycles. Digital PCR (dPCR), which is based on the amplification of single target DNA molecules in many separate reactions, is an attractive alternative to qPCR. Here we report a novel and easy method for multiplexing dPCR in picolitre droplets within emulsions-generated and read out in microfluidic devices-that takes advantage of both the very high numbers of reactions possible within emulsions (>10(6)) as well as the high likelihood that the amplification of only a single target DNA molecule will initiate within each droplet. By varying the concentration of different fluorogenic probes of the same color, it is possible to identify the different probes on the basis of fluorescence intensity. Adding multiple colors increases the number of possible reactions geometrically, rather than linearly as with qPCR. Accurate and precise copy numbers of up to sixteen per cell were measured using a model system. A 5-plex assay for spinal muscular atrophy was demonstrated with just two fluorophores to simultaneously measure the copy number of two genes (SMN1 and SMN2) and to genotype a single nucleotide polymorphism (c.815A>G, SMN1). Results of a pilot study with SMA patients are presented. 相似文献
77.
Quantitative and sensitive detection of rare mutations using droplet-based microfluidics 总被引:2,自引:0,他引:2
Pekin D Skhiri Y Baret JC Le Corre D Mazutis L Salem CB Millot F El Harrak A Hutchison JB Larson JW Link DR Laurent-Puig P Griffiths AD Taly V 《Lab on a chip》2011,11(13):2156-2166
Somatic mutations within tumoral DNA can be used as highly specific biomarkers to distinguish cancer cells from their normal counterparts. These DNA biomarkers are potentially useful for the diagnosis, prognosis, treatment and follow-up of patients. In order to have the required sensitivity and specificity to detect rare tumoral DNA in stool, blood, lymph and other patient samples, a simple, sensitive and quantitative procedure to measure the ratio of mutant to wild-type genes is required. However, techniques such as dual probe TaqMan(?) assays and pyrosequencing, while quantitative, cannot detect less than ~1% mutant genes in a background of non-mutated DNA from normal cells. Here we describe a procedure allowing the highly sensitive detection of mutated DNA in a quantitative manner within complex mixtures of DNA. The method is based on using a droplet-based microfluidic system to perform digital PCR in millions of picolitre droplets. Genomic DNA (gDNA) is compartmentalized in droplets at a concentration of less than one genome equivalent per droplet together with two TaqMan(?) probes, one specific for the mutant and the other for the wild-type DNA, which generate green and red fluorescent signals, respectively. After thermocycling, the ratio of mutant to wild-type genes is determined by counting the ratio of green to red droplets. We demonstrate the accurate and sensitive quantification of mutated KRAS oncogene in gDNA. The technique enabled the determination of mutant allelic specific imbalance (MASI) in several cancer cell-lines and the precise quantification of a mutated KRAS gene in the presence of a 200,000-fold excess of unmutated KRAS genes. The sensitivity is only limited by the number of droplets analyzed. Furthermore, by one-to-one fusion of drops containing gDNA with any one of seven different types of droplets, each containing a TaqMan(?) probe specific for a different KRAS mutation, or wild-type KRAS, and an optical code, it was possible to screen the six common mutations in KRAS codon 12 in parallel in a single experiment. 相似文献
78.
Peter Vorobieff C. Randall Truman Adam M. Ragheb Gregory S. Elliott Julia K. Laystrom-Woodard Darren M. King David L. Carroll Wayne C. Solomon 《Experiments in fluids》2011,51(3):711-722
We present quantitative analysis of image sequences of multi-stream injection nozzle flows with several different injection
geometries in an experiment simulating mixing in a chemical oxygen-iodine laser. To visualize mixing, image sequences were
acquired with planar laser-induced fluorescence (PLIF) in iodine that was injected into the main flow. The injection nozzle
consisted of a slot, ejector, and injector block, with rows of ejector and injector holes along the slot length. The ejector
flow exits in an underexpanded state so that upon expanding it forces the slot and injector flows together to enhance mixing.
For this study, the diameter and geometry of ejector holes were varied to assess their effect on mixing. Two configurations
of ejector holes were used, each with two different diameters for a total of four cases with data collected at downstream
stations. We carry out a quantitative mixing analysis for these configurations, using two methods to quantify the mixing.
The first method considers the statistics of the PLIF image intensity histograms, which are bimodal for poorly-mixed flows
and have a single peak in well-mixed flows. The second method quantifies the properties of the mixing interface. Our analysis
shows that two injection schemes significantly enhance mixing by stretching the mixing interface. 相似文献
79.
Hugh Rieley Daniel J. Colby Darren P. McMurray Stuart M. Reeman 《Surface science》1997,390(1-3):243-249
The effects of physisorption and two-dimensional ordering on the photochemistry of N2O4 were investigated. Ordered monolayers were prepared by adsorption of NO2 at 100 K on a water-ice surface. Irradiation with a continuous light source in the wavelength region 300–400 nm or with pulsed laser radiation at 355 nm resulted in exclusive desorption of NO2. This desorption was induced by electronic absorption directly in the adsorbate via a transition corresponding to the (
)1B2u←(
)1Ag transition in N2O4, as in the gas phase. However, the subsequent dynamics in the excited state were markedly different from the gas-phase counterpart. Time-of-flight mass spectrometry of NO2 photodesorbed at 355 nm revealed a most probable fragment translational energy of ca. 17 meV; and the angular distribution of the nascent NO2 was peaked sharply in a direction around 10° from the normal. It is apparent that, despite the weak interaction with the substrate, significant energy transfer occurs in the ordered physisorbed monolayer to yield nascent NO2 with very low translational energy and a constrained angle of escape which is consistent with a high degree of adsorbate order and alignment. 相似文献
80.