{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T03:06:41Z","timestamp":1791256001195,"version":"4.1.0"},"reference-count":46,"publisher":"National Academy of Sciences","issue":"18","content-domain":{"domain":["www.pnas.org"],"crossmark-restriction":true},"short-container-title":["Proc. Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[2011,5,3]]},"abstract":"<jats:p>\n                    The posttranslational modification of therapeutic proteins with terminal sialic acids is one means of improving their circulating half-life, thereby improving their efficiency. We have developed a two-step in vitro enzymatic modification of glycoproteins, which has previously only been achieved by chemical means [Gregoriadis G, Jain S, Papaioannou I, Laing P (2005)\n                    <jats:italic>Int J Pharm<\/jats:italic>\n                    300:125\u2013130). This two-step procedure uses the\n                    <jats:italic>Campylobacter jejuni<\/jats:italic>\n                    Cst-II \u03b12,8-sialyltransferase to provide a primer on N-linked glycans, followed by polysialylation using the\n                    <jats:italic>Neisseria meningitidis<\/jats:italic>\n                    \u03b12,8-polysialyltransferase. Here, we have demonstrated the ability of this system to modify three glycoproteins with varying N-linked glycan compositions: the human therapeutic proteins alpha-1-antitrypsin (A1AT) and factor IX, as well as bovine fetuin. The chain length of the polysialic acid addition was optimized by controlling reaction conditions. After demonstrating the ability of this system to modify a variety of proteins, the effect of polysialylation on the activity and serum half-life of A1AT was examined. The polysialylation of A1AT did not adversely affect its in vitro inhibition activity against human neutrophil elastase. The polysialylation of A1AT resulted in a significantly improved pharmacokinetic profile when the modified proteins were injected into CD-1 mice. Together, these results suggest that polysialylated A1AT may be useful for improved augmentation therapy for patients with a deficiency in this protein and that this modification may be applied to other therapeutic proteins.\n                  <\/jats:p>","DOI":"10.1073\/pnas.1019266108","type":"journal-article","created":{"date-parts":[[2011,4,19]],"date-time":"2011-04-19T00:19:39Z","timestamp":1303172379000},"page":"7397-7402","update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":104,"title":["Site-specific enzymatic polysialylation of therapeutic proteins using bacterial enzymes"],"prefix":"10.1073","volume":"108","author":[{"given":"Theresa","family":"Lindhout","sequence":"first","affiliation":[{"name":"Department of Molecular and Cellular Biology, University of Guelph, Guelph, ON, Canada N1G 2W1; and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Umar","family":"Iqbal","sequence":"additional","affiliation":[{"name":"Institute for Biological Sciences, National Research Council Canada, 100 Sussex Drive, Ottawa, ON, Canada K1A 0R6"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lisa M.","family":"Willis","sequence":"additional","affiliation":[{"name":"Department of Molecular and Cellular Biology, University of Guelph, Guelph, ON, Canada N1G 2W1; and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anne N.","family":"Reid","sequence":"additional","affiliation":[{"name":"Department of Molecular and Cellular Biology, University of Guelph, Guelph, ON, Canada N1G 2W1; and"},{"name":"Institute for Biological Sciences, National Research Council Canada, 100 Sussex Drive, Ottawa, ON, Canada K1A 0R6"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jianjun","family":"Li","sequence":"additional","affiliation":[{"name":"Institute for Biological Sciences, National Research Council Canada, 100 Sussex Drive, Ottawa, ON, Canada K1A 0R6"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xin","family":"Liu","sequence":"additional","affiliation":[{"name":"Institute for Biological Sciences, National Research Council Canada, 100 Sussex Drive, Ottawa, ON, Canada K1A 0R6"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria","family":"Moreno","sequence":"additional","affiliation":[{"name":"Institute for Biological Sciences, National Research Council Canada, 100 Sussex Drive, Ottawa, ON, Canada K1A 0R6"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Warren W.","family":"Wakarchuk","sequence":"additional","affiliation":[{"name":"Department of Molecular and Cellular Biology, University of Guelph, Guelph, ON, Canada N1G 2W1; and"},{"name":"Institute for Biological Sciences, National Research Council Canada, 100 Sussex Drive, Ottawa, ON, Canada K1A 0R6"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"341","published-online":{"date-parts":[[2011,4,18]]},"reference":[{"key":"e_1_3_3_1_2","doi-asserted-by":"publisher","DOI":"10.1038\/nbt1252"},{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.1038\/nrd1033"},{"key":"e_1_3_3_3_2","doi-asserted-by":"publisher","DOI":"10.2165\/00063030-200822050-00004"},{"key":"e_1_3_3_4_2","doi-asserted-by":"publisher","DOI":"10.1093\/glycob\/cwl004"},{"key":"e_1_3_3_5_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0169-409X(03)00108-X"},{"key":"e_1_3_3_6_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ijpharm.2005.06.007"},{"key":"e_1_3_3_7_2","doi-asserted-by":"publisher","DOI":"10.1038\/nrn2285"},{"key":"e_1_3_3_8_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M300458200"},{"key":"e_1_3_3_9_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0912103107"},{"key":"e_1_3_3_10_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0167-4838(97)00056-3"},{"key":"e_1_3_3_11_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0304-4165(03)00116-8"},{"key":"e_1_3_3_12_2","doi-asserted-by":"publisher","DOI":"10.1021\/bc8005122"},{"key":"e_1_3_3_13_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.271.4.1829"},{"key":"e_1_3_3_14_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M110.140038"},{"key":"e_1_3_3_15_2","doi-asserted-by":"publisher","DOI":"10.1093\/glycob\/cwm126"},{"key":"e_1_3_3_16_2","doi-asserted-by":"publisher","DOI":"10.2147\/COPD.S8577"},{"key":"e_1_3_3_17_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.rmed.2010.01.016"},{"key":"e_1_3_3_18_2","doi-asserted-by":"publisher","DOI":"10.1164\/ajrccm\/140.5.1494"},{"key":"e_1_3_3_19_2","first-page":"193","article-title":"Safety and efficacy of alpha-1-antitrypsin augmentation therapy in the treatment of patients with alpha-1-antitrypsin deficiency","volume":"3","author":"Petrache I","year":"2009","unstructured":"I Petrache, J Hajjar, M Campos, Safety and efficacy of alpha-1-antitrypsin augmentation therapy in the treatment of patients with alpha-1-antitrypsin deficiency. 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