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微晶纤维素在小鼠中递送重组蛋白基抗丹毒抗原的应用

Microcrystalline Cellulose for Delivery of Recombinant Protein-Based Antigen against Erysipelas in Mice.

机构信息

Biotechnology Process Engineering Center, KRIBB, Cheongju 363-883, Republic of Korea.

Department of Chemical and Biomolecular Engineering, KAIST, Daejeon 305-701, Republic of Korea.

出版信息

Biomed Res Int. 2018 Jun 11;2018:7670505. doi: 10.1155/2018/7670505. eCollection 2018.

Abstract

The study describes the development of a vaccine using microcrystalline cellulose (Avicel PH-101) as a delivery carrier of recombinant protein-based antigen against erysipelas. Recombinant SpaA, surface protective protein, from a gram-positive pathogen was fused to a cellulose-binding domain (CBD) from endoglucanase II through a S3N10 peptide. The fusion protein (CBD-SpaA) was expressed in and was subsequently bound to Avicel PH-101. The antigenicity of CBD-SpaA bound to the Avicel was evaluated by enzyme-linked immunosorbent (ELISA) and confocal laser scanning microscope (CLSM) assays. For the examination of its immunogenicity, groups of mice were immunized with different constructs (soluble CBD-SpaA, Avicel coated with CBD-SpaA, whole bacterin of (positive control), and PBS (negative control)). In two weeks after immunization, mice were challenged with 1x10 CFU of and Avicel coated with CBD-SpaA induced protective immunity in mice. In conclusion, this study demonstrates the feasibility of microcrystalline cellulose as the delivery system of recombinant protein subunit vaccine against infection in mice.

摘要

该研究描述了一种使用微晶纤维素(Avicel PH-101)作为载体递呈重组蛋白抗原的疫苗的研制,该抗原针对丹毒。将来自革兰氏阳性病原体的表面保护性蛋白重组 SpaA 通过 S3N10 肽融合到内切葡聚糖酶 II 的纤维素结合结构域(CBD)上。融合蛋白(CBD-SpaA)在 中表达,随后与 Avicel PH-101 结合。通过酶联免疫吸附(ELISA)和共聚焦激光扫描显微镜(CLSM)实验评估 CBD-SpaA 与 Avicel 结合的抗原性。为了检查其免疫原性,用不同的构建体(可溶性 CBD-SpaA、涂有 CBD-SpaA 的 Avicel、(阳性对照)的全菌和 PBS(阴性对照))对小鼠进行免疫。免疫后两周,用 1x10 CFU 的 对小鼠进行攻毒,涂有 CBD-SpaA 的 Avicel 诱导了小鼠的保护性免疫。总之,该研究证明了微晶纤维素作为针对 感染的重组蛋白亚单位疫苗递呈系统在小鼠中的可行性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3a39/6016178/e06f69b5e6b4/BMRI2018-7670505.001.jpg

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