caa a22 4500 445325151 CHVBK 20180317142718.0 cr unu---uuuuu 170323e20111101xx s 000 0 eng 10.1007/s10858-011-9563-8 doi (NATIONALLICENCE)springer-10.1007/s10858-011-9563-8 The use of 1H-31P GHMBC and covariance NMR to unambiguously determine phosphate ester linkages in complex polysaccharide mixtures [Elektronische Daten] [Edward Zartler, Gary Martin] Poly- and oligo-saccharides are commonly employed as antigens in many vaccines. These antigens contain phosphoester structural elements that are crucial to the antigenicity, and hence the effectiveness of the vaccine. Nuclear Magnetic Resonance (NMR) is a powerful tool for the site-specific identification of phosphoesters in saccharides. We describe here two advances in the characterization of phosphoesters in saccharides: (1) the use of 1H-31P GHMBC to determine the site-specific identity of phosphoester moieties in heterogeneous mixtures and (2) the use of Unsymmetrical/Generalized Indirect Covariance (U/GIC) to calculate a carbon-phosphorus 2D spectrum. The sensitivity of the 1H-31P GHMBC is far greater than the "standard” 1H-31P GHSQC and allows long-range 3-5JHP couplings to be readily detected. This is the first example to be reported of using U/GIC to calculate a carbon-phosphorus spectrum. The U/GIC processing affords, in many cases, a fivefold to tenfold or greater increase in signal-to-noise ratios in the calculated spectrum. When coupled with the high sensitivity of 1H-31P HMBC, U/GIC processing allows the complete and unambiguous assignments of phosphoester moieties present in heterogeneous samples at levels of ~5% (or less) of the total sample, expanding the breadth of samples that NMR can be used to analyze. This new analytical technique is generally applicable to any NMR-observable phosphoester. Springer Science+Business Media B.V., 2011 Phosphorus nationallicence Unsymmetrical indirect covariance nationallicence Generalized indirect covariance nationallicence Multiple bond coupling nationallicence Carbon-phosphorus spectrum nationallicence αRha : l-α-Rhamnose nationallicence βGlc : β-d-Glucose nationallicence β-ManNAc : β-2-Acetamido-mannose nationallicence β-GlcNAc : β-2-Acetamido-glucose nationallicence α-GlcNAc : α-2-Acetamido-glucose nationallicence NMR : Nuclear magnetic resonance spectroscopy nationallicence GHMBC : Gradient heteronuclear multiple bond correlation nationallicence GHMQC : Gradient heteronuclear multiple quantum correlation nationallicence GHSQC : Gradient heteronuclear single quantum correlation nationallicence PC : Phosphocholine nationallicence U/GIC : Unsymmetrical/generalized indirect covariance nationallicence MS : Mass spectrometry nationallicence Zartler Edward Vaccine Analytical Development, Merck Research Labs, Merck & Co., 19486, West Point, PA, USA aut Martin Gary Structure Elucidation Group, Merck Research Labs, Merck & Co., 07901, Summit, NJ, USA aut Journal of Biomolecular NMR Springer Netherlands 51/3(2011-11-01), 357-367 0925-2738 51:3<357 2011 51 10858 https://doi.org/10.1007/s10858-011-9563-8 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10858-011-9563-8 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Zartler Edward Vaccine Analytical Development, Merck Research Labs, Merck & Co., 19486, West Point, PA, USA aut NATIONALLICENCE 700 1- Martin Gary Structure Elucidation Group, Merck Research Labs, Merck & Co., 07901, Summit, NJ, USA aut NATIONALLICENCE 773 0- Journal of Biomolecular NMR Springer Netherlands 51/3(2011-11-01), 357-367 0925-2738 51:3<357 2011 51 10858 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer