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31.
Bing Gong Erik Hoyt Heather Lynaugh Irina Burnina Renee Moore Alissa Thompson Huijuan Li 《Analytical and bioanalytical chemistry》2013,405(17):5825-5831
N-Linked glycosylation is a major protein modification involved in many essential cellular functions. Methods capable of quantitative glycan analysis are highly valuable and have been actively pursued. Here we describe a novel N-glycosylamine-based strategy for isotopic labeling of N-linked glycans for quantitative analysis by use of mass spectrometry (MS). This strategy relies on the primary amine group on the reducing end of freshly released N-linked glycans for labeling, and eliminates the need for the harsh labeling reaction conditions and/or tedious cleanup procedures required by existing methods. By using NHS-ester amine chemistry we used this strategy to label N-linked glycans from a monoclonal antibody with commercially available tandem mass tags (TMT). Only duplex experiments can be performed with currently available TMT reagents, because quantification is based on the intensity of intact labeled glycans. Under mild reaction conditions, greater than 95 % derivatization was achieved in 30 min and the labeled glycans, when kept at ?20 °C, were stable for more than 10 days. By performing glycan release, TMT labeling, and LC–MS analysis continuously in a single volatile aqueous buffer without cleanup steps, we were able to complete the entire analysis in less than 2 h. Quantification was highly accurate and the dynamic range was large. Compared with previously established methods, N-glycosylamine-mediated labeling has the advantages of experimental simplicity, efficient labeling, and preserving glycan integrity.
Principle of N-Glycosylamine-mediated isotope labeling for mass spectrometry-based quantitative analysis of N-linked glycans 相似文献
32.
Kim HJ Deng J Lalli JH Riffle JS Viers BD Esker AR 《Langmuir : the ACS journal of surfaces and colloids》2005,21(5):1908-1916
Mixtures of a polyhedral oligomeric silsesquioxane, trisilanolisobutyl-POSS, and a polar silicone, poly(dimethyl-co-methylvinyl-co-methyl, 2-diphenyl phosphine oxide ethyl) siloxane (PDMS-PO), spread as Langmuir monolayers at the air/water interface are used to examine the surface phase behavior and aggregation of trisilanolisobutyl-POSS as a function of silicone composition. Analyses of the surface pressure-area per monomer (Pi-A) isotherms in terms of the collapse pressures and excess Gibbs free energies of mixing indicate the monolayers form slightly negative deviation mixtures. Direct observations of surface morphology with Brewster angle microscopy in the collapsed regime reveal that the governing factor for aggregation is the collapse Pi of the component with a stronger affinity for water. In trisilanolisobutyl-POSS/PDMS-PO blends, POSS aggregates as discrete domains and does not coalesce into larger aggregates or networklike structures for <80 wt % POSS, a feature that is vastly different from a previous study of POSS blended with regular poly(dimethylsiloxane). 相似文献
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A detailed study of the dialkylation of dianions derived from dihydroisoindigo 1 with enantiopure ditriflate 2 is reported. The LHMDS-mediated process has been optimized to give C(2)-symmetric product 3 with high selectivity (C(2) selectivity 3:5 = 100:1; C(2):C(1) selectivity = 8:1). Stereoselection in the C(2) manifold is determined in both the bimolecular and intramolecular alkylation steps. 相似文献
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Hoyt CW Barber ZW Oates CW Fortier TM Diddams SA Hollberg L 《Physical review letters》2005,95(8):083003
We report the direct excitation of the highly forbidden (6s2) 1S0 <--> (6s6p) 3P0 optical transition in two odd isotopes of neutral ytterbium. As the excitation laser frequency is scanned, absorption is detected by monitoring the depletion from an atomic cloud at approximately 70 microK in a magneto-optical trap. The measured frequency in 171Yb (F=1/2) is 518,295,836,591.6 +/- 4.4 kHz. The measured frequency in 173Yb (F=5/2) is 518,294,576,847.6 +/- 4.4 kHz. Measurements are made with a femtosecond-laser frequency comb calibrated by the National Institute of Standards and Technology cesium fountain clock and represent nearly a 10(6)-fold reduction in uncertainty. The natural linewidth of these J=0 to J=0 transitions is calculated to be approximately 10 mHz, making them well suited to support a new generation of optical atomic clocks based on confinement in an optical lattice. 相似文献
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Crystal Hoyt 《Israel Journal of Mathematics》2012,192(1):251-280
We classify good ?-gradings of basic Lie superalgebras over an algebraically closed field $\mathbb{F}$ of characteristic zero. Good ?-gradings are used in quantum Hamiltonian reduction for affine Lie superalgebras, where they play a role in the construction of super W-algebras. We also describe the centralizer of a nilpotent even element and of an $\mathfrak{s}\mathfrak{l}_2$ -triple in $\mathfrak{g}\mathfrak{l}\left( {\left. m \right|n} \right)$ and $\mathfrak{o}\mathfrak{s}\mathfrak{p}\left( {\left. m \right|2n} \right)$ . 相似文献