title: Expression of CTB-10×rolGLP-1 in E. coli and its therapeutic effect on type 2 diabetes.
abstract: Glucagon-like peptide-1 (GLP-1) is a short peptide that can significantly reduce blood glucose level. Recombination oral long-acting glucagon-like peptide-1 (rolGLP-1), is a GLP-1 analog generated from site-specific mutation of GLP-1. CTB is a non-toxic portion of the cholera toxin and an ideal protein antigen carrier. In this study, we firstly constructed a vector pET-22b (+)-CTB-10×rolGLP-1 to express a fusion protein composed of CTB and ten tandem repeated rolGLP-1 in BL21 (DE3) line of E. coli. The CTB-10×rolGLP-1 was expressed efficiently in the inclusion bodies. The expression product was analyzed by SDS-PAGE electrophoresis and Western blotting. The inclusion bodies were then denatured, refolded and purified by ion exchange chromatography to obtain a high-purity CTB- 10×rolGLP-1. The therapeutic effect of CTB-10×rolGLP-1 was assessed in comparison with 10×rolGLP-1 alone by daily oral-gavage administration up to 10 days in streptozotocin-induced type 2 diabetic mice. The results showed that the level of blood glucose was reduced more effectively and the oral glucose tolerance of mice was improved more significantly with the administration of CTB-10×rolGLP-1. Our results provided a potentially promising oral biological drug for the treatment of type 2 diabetes.
authors: Peipei
authors: P
authors: Tu
authors: Zhihua
authors: Z
authors: Ma
authors: Haisong
authors: H
authors: Wang
authors: Baicheng
authors: B
authors: Ma
authors: Xiaodan
authors: X
authors: Li
authors: Huikun
authors: H
authors: Duan
authors: Pingzhe
authors: P
authors: Jiang
authors: Miao
authors: M
authors: Li
authors: Ri
authors: R
authors: Wu
authors: Jianhong
authors: J
authors: Zhu
authors: Department of Preventive Medicine, Wenzhou Medical University, Wenzhou, Zhejiang 325035, China; Key Laboratory for Bioactive Materials of the Ministry of Education, Institute of Molecular Biology, College of Life Science, Nankai University, Weijin Road 94, Nankai District, 300071, Tianjin, China. jianhong.zhu@gmail.com.
authors: Minggang
authors: M
authors: Li
mesh_terms: D000284:Administration, Oral; D000818:Animals; D001786:Blood Glucose / Q000032:analysis; D002772:Cholera Toxin / Q000008:administration & dosage* / Q000235:genetics; D003924:Diabetes Mellitus, Type 2 / Q000097:blood* / Q000175:diagnosis / Q000188:drug therapy*; D004338:Drug Combinations; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D052216:Glucagon-Like Peptide 1 / Q000008:administration & dosage* / Q000235:genetics; D007004:Hypoglycemic Agents / Q000008:administration & dosage; D008297:Male; D051379:Mice; D015202:Protein Engineering / Q000379:methods; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage / Q000235:genetics / Q000378:metabolism; D016896:Treatment Outcome
chemical_list: D001786:Blood Glucose; D004338:Drug Combinations; D007004:Hypoglycemic Agents; D011993:Recombinant Fusion Proteins; D052216:Glucagon-Like Peptide 1; D002772:Cholera Toxin