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

CHEN Jiaqi, ZHANG Qi, PAN Pengwei, BAI Fang, GENG Peng, GAO Zhihui, BAI Gang

doi:10.1271/bbb.80742

Bibliographic Information

Other Title

Development, Characterization, and Evaluation of a Fusion Protein of a Novel Glucagon-Like Peptide-1 (GLP-1) Analog and Human Serum Albumin in<i>Pichia pastoris</i>

Published: 2009

DOI

10.1271/bbb.80742

Publisher: Japan Society for Bioscience, Biotechnology, and Agrochemistry

Search this article

Description

Glucagon-like peptide-1 (GLP-1) has considerable potential as a possible therapeutic agent for type-2 diabetes. Unfortunately, this glucoincretin is short lived due to degradation by dipeptidyl-peptidase IV and rapid clearance by renal filtration. In this study, we attempted to extend GLP-1 action through the attachment of a lysine residue at the N-terminal of GLP-1 (named KGLP-1), and to make a fusion protein with human serum albumin (HSA) in Pichia pastoris. The protein, designated KGLP-1/HSA, was purified by an immunomagnetic separation technique. High performance liquid chromatography (HPLC) showed that the purified protein had an overall purity of 92.0%, and matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS) confirmed the expected molecular mass of 70,297.8 Da. Additionally, the N-terminal sequence of KGLP-1/HSA was confirmed by N-terminal sequencing. The stability and biological activity of KGLP-1/HSA were then evaluated in vitro and in vivo. The findings indicated that fusion KGLP-1/HSA preserved the action of native GLP-1, and the active duration was greatly prolonged.

Journal

Bioscience, Biotechnology, and Biochemistry

Bioscience, Biotechnology, and Biochemistry 73 (3), 688-694, 2009

Japan Society for Bioscience, Biotechnology, and Agrochemistry

Keywords

Details 詳細情報について

Export

Report a problem