{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,26]],"date-time":"2025-10-26T14:26:17Z","timestamp":1761488777428,"version":"build-2065373602"},"reference-count":44,"publisher":"Wiley","issue":"10","license":[{"start":{"date-parts":[[2011,10,5]],"date-time":"2011-10-05T00:00:00Z","timestamp":1317772800000},"content-version":"vor","delay-in-days":4,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["J. Mass. Spectrom."],"published-print":{"date-parts":[[2011,10]]},"abstract":"<jats:p>We present herein an efficient mass spectrometric method for the localization of the glycation sites of a model neoglycoconjugate vaccine formed by a construct of the tetrasaccharide side chain of the <jats:italic>Bacillus anthracis<\/jats:italic> exosporium and the protein carrier bovine serum albumin. The glycoconjugate was digested with both trypsin and GluC V8 endoproteinases, and the digests were then analyzed by MALDI\u2010TOF\/TOF\u2010CID\u2010MS\/MS and nano\u2010LC\u2010ESI\u2010QqTOF\u2010CID\u2010MS\/MS. The sequences of the unknown peptides analyzed by MALDI\u2010TOF\/TOF\u2010CID\u2010MS\/MS, following digestion with the GluC V8 endoproteinase, allowed us to recognize three glycopeptides whose glycation occupancies were, respectively, on Lys 235, Lys 420, and Lys 498. Similarly, the same analysis was performed on the tryptic digests, which permitted us to recognize two glycation sites on Lys 100 and Lys 374. In addition, we have also used LC\u2010ESI\u2010QqTOF\u2010CID\u2010MS\/MS analysis for the identification of the tryptic digests. However, this analysis identified a higher number of glycopeptides than would be expected from a glycoconjugate composed of a carbohydrate\u2013protein ratio of 5.4:1, which would have resulted in glycation occupancies of 18 specific sites. This discrepancy was due to the large number of glycoforms formed during the synthetic carbohydrate\u2013spacer\u2013carrier protein conjugation. Likewise, the LC\u2010ESI\u2010QqTOF\u2010MS\/MS analysis of the GluC V8 digest also identified 17 different glycation sites on the synthetic glycoconjugate. Copyright \u00a9 2011 John Wiley &amp; Sons, Ltd.<\/jats:p>","DOI":"10.1002\/jms.1980","type":"journal-article","created":{"date-parts":[[2011,10,6]],"date-time":"2011-10-06T00:07:13Z","timestamp":1317859633000},"page":"993-1003","source":"Crossref","is-referenced-by-count":14,"title":["Determination of the glycation sites of <i>Bacillus anthracis<\/i> neoglycoconjugate vaccine by MALDI\u2010TOF\/TOF\u2010CID\u2010MS\/MS and LC\u2010ESI\u2010QqTOF\u2010tandem mass spectrometry"],"prefix":"10.1002","volume":"46","author":[{"given":"Farid","family":"Jahouh","sequence":"first","affiliation":[{"name":"Department of Chemistry Memorial University of Newfoundland  Saint John's NL Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shu\u2010jie","family":"Hou","sequence":"additional","affiliation":[{"name":"NIDDK, LBC, National Institutes of Health  Bethesda MD 20892\u20100815 USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Pavol","family":"Kov\u00e1\u010d","sequence":"additional","affiliation":[{"name":"NIDDK, LBC, National Institutes of Health  Bethesda MD 20892\u20100815 USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Joseph H.","family":"Banoub","sequence":"additional","affiliation":[{"name":"Department of Chemistry Memorial University of Newfoundland  Saint John's NL Canada"},{"name":"Science Branch, Special Projects Department of Fisheries and Oceans Canada  Saint John's NL A1C 5X1 Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2011,10,5]]},"reference":[{"doi-asserted-by":"publisher","key":"e_1_2_5_2_1","DOI":"10.1016\/j.carres.2003.07.008"},{"key":"e_1_2_5_3_1","first-page":"187","volume-title":"Handbook of Endotoxins, Chemistry of Endotoxins","author":"Rietschel E. 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