One-Step Purification of a Fusion Protein of Glucagon-Like Peptide-1 and Human Serum Albumin Expressed in Pichia pastoris by an Immunomagnetic Separation Technique

スポンサーリンク

概要

• 論文の詳細を見る

• Glucagon-like peptide-1 (GLP-1) has great therapeutic potential to treat diabetes type 2, mainly due to its unique glucose-dependent stimulation of insulin secretion profiles, but its clinical application is limited by its short half-life in vivo, which resultes from degradation by dipeptidyl peptidase IV and/or renal clearance. Developing long-acting GLP-1 analogs is therefore an important step toward using them therapeutically. In this study, the GLP-1/human serum albumin (HSA) fusion protein gene was cloned into the secretor type expression vector pPIC9K and subsequently expressed in Pichia pastoris. The expression quantity reached 58.5 mg/l in small-scale incubation. After optimization and characterization, the GLP-1/HSA fusion protein was successfully purified from the supernatant of the broth using immunomagnetic cellulose microspheres. HPLC showed that the purified GLP-1/HSA had an overall purity of 93.9%, and matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS) confirmed the fusion protein exhibited the expected molecular mass of 70 kDa. Furthermore, that analysis of in vivo activity indicated that GLP-1/HSA reduced the blood glucose level after intraperitoneal administration to Chinese Kunming mice in a dose-dependent manner, and the effects held significantly 4 h after administration. Overall, this study illustrates the development of a long-acting GLP-1/HSA fusion protein expressed in Pichia pastoris.

• 社団法人 日本農芸化学会の論文

著者

• ZHANG Qi

  College of Life Science and Pharmaceutical Engineering, Nanjing University of Technology

• Cao Yu

  Department Of Microbiology College Of Life Sciences Nankai University

• Bai Gang

  Department Of Microbiology College Of Life Sciences Nankai University

• YANG Wenbo

  Department of Microbiology, College of Life Sciences, Nankai University

• CHEN Jiaqi

  Department of Microbiology, College of Life Sciences, Nankai University

• GAO Zhihui

  Department of Microbiology, College of Life Sciences, Nankai University

• ZHU Yuanyuan

  College of Pharmaceutical Sciences, Nankai University

関連論文

• Cloning, Expression, and Identification of Genes Involved in the Conversion of DL-2-Amino-Δ^2-thiazoline-4-carboxylic Acid to L-Cysteine via S-Carbamyl-L-cysteine Pathway in Pseudomonas sp. TS1138
• Effects of strontium fructose 1,6-diphosphate on expression of apoptosis-related genes and oxidative stress in testes of diabetic rats
• One-step elimination of L-cysteine desulfhydrase from crude enzyme extracts of Pseudomonas sp. TS1138 using an immunomagnetic affinity matrix improves the enzymatic production of L-cysteine
• One-Step Purification of a Fusion Protein of Glucagon-Like Peptide-1 and Human Serum Albumin Expressed in Pichia pastoris by an Immunomagnetic Separation Technique
• Preparation and characterization of magnetic microspheres for the purification of interferon α-2b
• Pathogenic Cellulase Assay of Pine Wilt Disease and Immunological Localization
• One-Step Purification of a Fusion Protein of Glucagon-Like Peptide-1 and Human Serum Albumin Expressed in Pichia pastoris by an Immunomagnetic Separation Technique
• Development, Characterization, and Evaluation of a Fusion Protein of a Novel Glucagon-Like Peptide-1 (GLP-1) Analog and Human Serum Albumin in Pichia pastoris
• Use of an Enhanced Green Fluorescence Protein Linked to a Single Chain Fragment Variable Antibody to Localize Bursaphelenchus xylophilus Cellulase
• Single Dose Administration of L-Carnitine Improves Antioxidant Activities in Healthy Subjects
• Common Variation in With No-Lysine Kinase 1 (WNK1) and Blood Pressure Responses to Dietary Sodium or Potassium Interventions : Family-Based Association Study

もっと見る 閉じる

スポンサーリンク