{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,4]],"date-time":"2026-08-04T13:55:39Z","timestamp":1785851739487,"version":"3.56.0"},"reference-count":60,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2012,10,18]],"date-time":"2012-10-18T00:00:00Z","timestamp":1350518400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Plant Biotechnology Journal"],"published-print":{"date-parts":[[2013,1]]},"abstract":"<jats:title>Summary<\/jats:title><jats:p>Glucagon\u2010like peptide (<jats:styled-content style=\"fixed-case\">GLP<\/jats:styled-content>\u20101) increases insulin secretion but is rapidly degraded (half\u2010life: 2\u00a0min in circulation). <jats:styled-content style=\"fixed-case\">GLP<\/jats:styled-content>\u20101 analogue, exenatide (<jats:styled-content style=\"fixed-case\">B<\/jats:styled-content>yetta) has a longer half\u2010life (3.3\u20134\u00a0h) with potent insulinotropic effects but requires cold storage, daily abdominal injections with short shelf life. Because patients with diabetes take &gt;60\u00a0000 injections in their life time, alternative delivery methods are highly desired. Exenatide is ideal for oral delivery because insulinotropism is glucose dependent, with reduced risk of hypoglycaemia even at higher doses. Therefore, exendin\u20104 (<jats:styled-content style=\"fixed-case\">EX4<\/jats:styled-content>) was expressed as a cholera toxin <jats:styled-content style=\"fixed-case\">B<\/jats:styled-content> subunit (<jats:styled-content style=\"fixed-case\">CTB<\/jats:styled-content>)\u2013fusion protein in tobacco chloroplasts to facilitate bioencapsulation within plant cells and transmucosal delivery in the gut via <jats:styled-content style=\"fixed-case\">GM<\/jats:styled-content>1 receptors present in the intestinal epithelium. The transgene integration was confirmed by <jats:styled-content style=\"fixed-case\">PCR<\/jats:styled-content> and <jats:styled-content style=\"fixed-case\">S<\/jats:styled-content>outhern blot analysis. Expression level of <jats:styled-content style=\"fixed-case\">CTB<\/jats:styled-content>\u2010<jats:styled-content style=\"fixed-case\">EX<\/jats:styled-content>4 reached up to 14.3% of total leaf protein (<jats:styled-content style=\"fixed-case\">TLP<\/jats:styled-content>). Lyophilization of leaf material increased therapeutic protein concentration by 12\u2010 to 24\u2010fold, extended their shelf life up to 15\u00a0months when stored at room temperature and eliminated microbes present in fresh leaves. The pentameric structure, disulphide bonds and functionality of <jats:styled-content style=\"fixed-case\">CTB<\/jats:styled-content>\u2010<jats:styled-content style=\"fixed-case\">EX<\/jats:styled-content>4 were well preserved in lyophilized materials. Chloroplast\u2010derived <jats:styled-content style=\"fixed-case\">CTB<\/jats:styled-content>\u2010<jats:styled-content style=\"fixed-case\">EX<\/jats:styled-content>4 showed increased insulin secretion similar to the commercial <jats:styled-content style=\"fixed-case\">EX<\/jats:styled-content>4 in beta\u2010<jats:styled-content style=\"fixed-case\">TC<\/jats:styled-content>6, a mouse pancreatic cell line. Even when 5000\u2010fold excess dose of <jats:styled-content style=\"fixed-case\">CTB<\/jats:styled-content>\u2010<jats:styled-content style=\"fixed-case\">EX<\/jats:styled-content>4 was orally delivered, it stimulated insulin secretion similar to the intraperitoneal injection of commercial <jats:styled-content style=\"fixed-case\">EX<\/jats:styled-content>4 but did not cause hypoglycaemia in mice. Oral delivery of the bioencapsulated <jats:styled-content style=\"fixed-case\">EX<\/jats:styled-content>4 should eliminate injections, increase patient compliance\/convenience and significantly lower their cost.<\/jats:p>","DOI":"10.1111\/pbi.12008","type":"journal-article","created":{"date-parts":[[2012,10,18]],"date-time":"2012-10-18T19:24:11Z","timestamp":1350588251000},"page":"77-86","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":91,"title":["Oral delivery of bioencapsulated exendin\u20104 expressed in chloroplasts lowers blood glucose level in mice and stimulates insulin secretion in beta\u2010<scp>TC<\/scp>6 cells"],"prefix":"10.1111","volume":"11","author":[{"given":"Kwang\u2010Chul","family":"Kwon","sequence":"first","affiliation":[{"name":"Department of Molecular Biology and Microbiology College of Medicine University of Central Florida Orlando FL USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ramya","family":"Nityanandam","sequence":"additional","affiliation":[{"name":"Department of Molecular Biology and Microbiology College of Medicine University of Central Florida Orlando FL USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"James S.","family":"New","sequence":"additional","affiliation":[{"name":"Department of Molecular Biology and Microbiology College of Medicine University of Central Florida Orlando FL USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Henry","family":"Daniell","sequence":"additional","affiliation":[{"name":"Department of Molecular Biology and Microbiology College of Medicine University of Central Florida Orlando FL USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2012,10,18]]},"reference":[{"key":"e_1_2_6_2_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1467-7652.2007.00258.x"},{"key":"e_1_2_6_3_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1163420"},{"key":"e_1_2_6_4_1","doi-asserted-by":"publisher","DOI":"10.1053\/j.gastro.2007.03.054"},{"key":"e_1_2_6_5_1","doi-asserted-by":"publisher","DOI":"10.2337\/diabetes.53.9.2492"},{"key":"e_1_2_6_6_1","doi-asserted-by":"publisher","DOI":"10.1126\/scitranslmed.3002621"},{"key":"e_1_2_6_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.tips.2010.06.003"},{"key":"e_1_2_6_8_1","first-page":"281","article-title":"Exenatide (Byetta) as a novel treatment option for type 2 diabetes mellitus","volume":"19","author":"Bond A.","year":"2006","journal-title":"Proc. (Bayl. Univ. Med. Cent.)"},{"key":"e_1_2_6_9_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1467-7652.2010.00582.x"},{"key":"e_1_2_6_10_1","volume-title":"National Diabetes Fact Sheet: National Estimates and General Information on Diabetes and Prediabetes in the United States, 2011","author":"Centers for Disease Control and Prevention","year":"2011"},{"key":"e_1_2_6_11_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ddmec.2005.08.013"},{"key":"e_1_2_6_12_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0702219104"},{"key":"e_1_2_6_13_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.tplants.2009.09.009"},{"key":"e_1_2_6_14_1","doi-asserted-by":"publisher","DOI":"10.1186\/1472-6750-9-33"},{"key":"e_1_2_6_15_1","doi-asserted-by":"publisher","DOI":"10.3949\/ccjm.76.s5.05"},{"key":"e_1_2_6_16_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1467-7652.2009.00479.x"},{"key":"e_1_2_6_17_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0167-0115(03)00120-4"},{"key":"e_1_2_6_18_1","unstructured":"FDA drug safety information(2009)Information for healthcare professionals: reports of altered kidney function in patients using Exenatide (marketed as Byetta). Available at:http:\/\/www.fda.gov\/Drugs\/DrugSafety\/PostmarketDrugSafetyInformationforPatientsandProviders\/DrugSafetyInformationforHeathcareProfessionals\/ucm188656.htm(accessed July 2012)."},{"key":"e_1_2_6_19_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1467-7652.2008.00338.x"},{"key":"e_1_2_6_20_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0022-2275(20)37914-1"},{"key":"e_1_2_6_21_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1467-7652.2011.00619.x"},{"key":"e_1_2_6_22_1","doi-asserted-by":"publisher","DOI":"10.1002\/9783527636730.ch3"},{"key":"e_1_2_6_23_1","first-page":"379","article-title":"Principles of preserving bacteria by freeze\u2010drying","volume":"26","author":"Heckly R.J.","year":"1985","journal-title":"Dev. Ind. Microbiol."},{"key":"e_1_2_6_24_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.mce.2004.04.017"},{"key":"e_1_2_6_25_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.72.7.2520"},{"key":"e_1_2_6_26_1","doi-asserted-by":"publisher","DOI":"10.1016\/0167-0115(95)00063-H"},{"key":"e_1_2_6_27_1","doi-asserted-by":"publisher","DOI":"10.2337\/diacare.28.5.1083"},{"key":"e_1_2_6_28_1","doi-asserted-by":"publisher","DOI":"10.1210\/endo.136.8.7628397"},{"key":"e_1_2_6_29_1","doi-asserted-by":"publisher","DOI":"10.1210\/jc.2002-021545"},{"key":"e_1_2_6_30_1","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.73.12.8266-8274.2005"},{"key":"e_1_2_6_31_1","doi-asserted-by":"publisher","DOI":"10.1097\/01.crd.0000223655.16253.e4"},{"key":"e_1_2_6_32_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1467-7652.2010.00538.x"},{"key":"e_1_2_6_33_1","doi-asserted-by":"publisher","DOI":"10.1096\/fj.05-5134fje"},{"key":"e_1_2_6_34_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrendo.2009.48"},{"key":"e_1_2_6_35_1","doi-asserted-by":"publisher","DOI":"10.1007\/978-94-007-2217-0_2"},{"key":"e_1_2_6_36_1","doi-asserted-by":"publisher","DOI":"10.1210\/me.2004-0350"},{"key":"e_1_2_6_37_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.coph.2008.06.002"},{"key":"e_1_2_6_38_1","doi-asserted-by":"publisher","DOI":"10.1093\/emboj\/18.3.595"},{"key":"e_1_2_6_39_1","volume-title":"Expression and functional evaluation of exendin 4 fused to cholera toxin B subunit in tobacco chloroplast to treat type 2 diabetes","author":"Nityanandam R.","year":"2011"},{"key":"e_1_2_6_40_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.imbio.2010.09.008"},{"key":"e_1_2_6_41_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.vaccine.2011.07.077"},{"key":"e_1_2_6_42_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.it.2008.09.006"},{"key":"e_1_2_6_43_1","doi-asserted-by":"publisher","DOI":"10.1002\/dmrr.646"},{"key":"e_1_2_6_44_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1467-7652.2007.00259.x"},{"key":"e_1_2_6_45_1","doi-asserted-by":"publisher","DOI":"10.1104\/pp.109.152017"},{"key":"e_1_2_6_46_1","doi-asserted-by":"publisher","DOI":"10.1086\/313951"},{"key":"e_1_2_6_47_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00018-008-7496-5"},{"key":"e_1_2_6_48_1","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2006.4"},{"key":"e_1_2_6_49_1","doi-asserted-by":"publisher","DOI":"10.1179\/acb.2003.58.6.001"},{"key":"e_1_2_6_50_1","doi-asserted-by":"publisher","DOI":"10.14573\/altex.2010.2.105"},{"key":"e_1_2_6_51_1","first-page":"451","article-title":"Animal models in type 2 diabetes research: an overview","volume":"125","author":"Srinivasan K.","year":"2007","journal-title":"Indian J. Med. Res."},{"key":"e_1_2_6_52_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2249.2004.02520.x"},{"key":"e_1_2_6_53_1","doi-asserted-by":"publisher","DOI":"10.2337\/diabetes.49.5.741"},{"key":"e_1_2_6_54_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-3083.2009.02321.x"},{"key":"e_1_2_6_55_1","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2007.522"},{"key":"e_1_2_6_56_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0912181107"},{"key":"e_1_2_6_57_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0958-1669(02)00284-7"},{"key":"e_1_2_6_58_1","doi-asserted-by":"publisher","DOI":"10.2337\/diacare.27.5.1047"},{"key":"e_1_2_6_59_1","doi-asserted-by":"publisher","DOI":"10.1136\/gut.8.6.618"},{"key":"e_1_2_6_60_1","doi-asserted-by":"publisher","DOI":"10.2337\/diabetes.48.5.1026"},{"key":"e_1_2_6_61_1","doi-asserted-by":"publisher","DOI":"10.4161\/hv.7.3.14207"}],"container-title":["Plant Biotechnology Journal"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1111%2Fpbi.12008","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/pbi.12008","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,11,22]],"date-time":"2023-11-22T11:57:23Z","timestamp":1700654243000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1111\/pbi.12008"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2012,10,18]]},"references-count":60,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2013,1]]}},"alternative-id":["10.1111\/pbi.12008"],"URL":"https:\/\/doi.org\/10.1111\/pbi.12008","archive":["Portico"],"relation":{},"ISSN":["1467-7644","1467-7652"],"issn-type":[{"value":"1467-7644","type":"print"},{"value":"1467-7652","type":"electronic"}],"subject":[],"published":{"date-parts":[[2012,10,18]]},"assertion":[{"value":"2012-07-18","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2012-09-18","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2012-10-18","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}