江苏农业学报2024,Vol.40Issue(1) :141-148.DOI:10.3969/j.issn.1000-4440.2024.01.015

包裹有CTA1-DD蛋白的OCS-DS纳米颗粒的制备及佐剂活性

Preparation and adjuvant activity of OCS-DS nanoparticles encapsulating CTA1-DD protein

秦竹 陈瑾 侯立婷 乔绪稳 李兰 杨利 杜露平 于晓明 张元鹏 郑其升
江苏农业学报2024,Vol.40Issue(1) :141-148.DOI:10.3969/j.issn.1000-4440.2024.01.015

包裹有CTA1-DD蛋白的OCS-DS纳米颗粒的制备及佐剂活性

Preparation and adjuvant activity of OCS-DS nanoparticles encapsulating CTA1-DD protein

秦竹 1陈瑾 1侯立婷 1乔绪稳 1李兰 1杨利 1杜露平 1于晓明 1张元鹏 1郑其升1
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作者信息

  • 1. 江苏省农业科学院动物免疫工程研究所/国家兽用生物制品工程技术研究中心/江苏省食品质量与安全重点实验室,江苏南京 210014;兽用生物制品国泰技术创新中心,江苏 泰州 225300
  • 折叠

摘要

重组CTA1-DD蛋白具有与完整CT分子相当的全身性和黏膜佐剂功能,但在复杂的生理环境中易被酶或酸降解.本研究以同样具有佐剂活性的O-羧甲基壳聚糖(OCS)和硫酸葡聚糖(DS)为载体,通过离子交联形成纳米颗粒,将CTA1-DD蛋白嵌入其中,使其得到稳定保护.包裹有CTA1-DD蛋白的OCS-DS纳米颗粒的粒径为50~150 nm,Zeta电位约-50 mV,质量浓度1.0 mg/ml的CTA1-DD蛋白投入制备的包裹有CTA1-DD蛋白的OCS-DS纳米颗粒载药率 25.33%,包封率 86.56%.体外模拟释放试验结果显示CTA1-DD蛋白可在 7d内缓慢释放.将CTA1-DD蛋白与PRV灭活抗原混合后,接种至小鼠鼻腔,结果表明,包裹有CTA1-DD蛋白的OCS-DS纳米颗粒能够同时诱导更高的血清IgG抗体和黏膜IgA抗体表达,证明了其作为黏膜佐剂的高效性.

Abstract

The recombinant CTA1-DD protein has systemic and mucosal adjuvant functions comparable to intact CT molecules.But it is easily degraded by enzymes or acids in complex physiological environments.In this study,O-carboxym-ethyl chitosan(OCS)and dextran sulfate(DS),which also had adjuvant activity,were used as carriers to form nanoparti-cles through ion crosslinking,and CTA1-DD protein was embedded in them for stable protection.The particle size of OCS-DS nanoparticles encapsulating CTA1-DD protein ranged from 50 nm to 150 nm,with a Zeta potential of approximately-50 mV.The drug loading rate of OCS-DS nanoparticles encapsulating CTA1-DD protein was 25.33%when 1.0 mg/ml CTA1-DD protein was put into prepara-tion,and the protein encapsulation rate was 86.56%.The simulated release tests in vitro showed that CTA1-DD pro-tein could be slowly released within seven days.The mice were inoculated intranasally with the mixture of CTA1-DD protein and PRV inactivated antigen.The results showed that OCS-DS nanoparticles encapsulated with CTA1-DD protein could simultaneously induce higher expression of serum IgG an-tibodies and mucosal IgA antibodies,which proved its high efficiency as mucosal adjuvant.

关键词

CTA1-DD蛋白/O-羧甲基壳聚糖/硫酸葡聚糖/纳米颗粒/佐剂活性

Key words

CTA1-DD protein/O-carboxymethyl chitosan/dextran sulfate/nanoparticles/adjuvant activity

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基金项目

江苏省农业科技自主创新基金项目(CX213135)

十四五重点研发专项(2022YFD1800800)

出版年

2024
江苏农业学报
江苏省农业科学院

江苏农业学报

CSTPCDCSCD北大核心
影响因子:1.093
ISSN:1000-4440
参考文献量26
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