期刊
VACCINE
卷 32, 期 39, 页码 5019-5026出版社
ELSEVIER SCI LTD
DOI: 10.1016/j.vaccine.2014.07.016
关键词
Cholera toxin B subunit; alpha-Helical coiled coil; Cartilage oligomeric matrix protein; Physicochemical stability; Rational design of vaccines
资金
- Cooperative Research Grant Program of the Institute of Tropical Medicine at Nagasaki University in Japan
- Bio-oriented Technology Research Advancement Institution of Japan
To create a physicochemically stable cholera toxin (CT) B subunit (CTB), it was fused to the five-stranded alpha-helical coiled-coil domain of cartilage oligomeric matrix protein (COMP). The chimeric fusion protein (CTB-COMP) was expressed in Pichia pastoris, predominantly as a pentamer, and retained its affinity for the monosialoganglioside GM1, a natural receptor of CT. The fusion protein displayed thermostability, tolerating the boiling temperature of water for 10 mm, whereas unfused CTB readily dissociated to its monomers and lost its affinity for GM1. The fusion protein also displayed resistance to strong acid at pHs as low as 0.1, and to the protein denaturant sodium dodecyl sulfate at concentrations up to 10%. Intranasal administration of the fusion protein to mice induced anti-B subunit serum IgG, even after the protein was boiled, whereas unfused CTB showed no thermostable mucosal immunogenicity. This study demonstrates that CTB fused to a pentameric alpha-helical coiled coil has a novel physicochemical phenotype, which may provide important insight into the molecular design of enterotoxin-B-subunit-based vaccines and vaccine delivery molecules. (C) 2014 Elsevier Ltd. All rights reserved.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据