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

作　　者：秦竹[1,2] 陈瑾 侯立婷[1,2] 乔绪稳 李兰[1,2] 杨利 杜露平[1,2] 于晓明 张元鹏[1,2] 郑其升 QIN Zhu;CHEN Jin;HOU Li-ting;QIAO Xu-wen;LI Lan;YANG Li;DU Lu-ping;YU Xiao-ming;ZHANG Yuan-peng;ZHENG Qi-sheng(Institute of Veterinary Immunology&Engineering,Jiangsu Academy of Agricultural Sciences/National Research Center of Engineering and Technology for Veterinary Biologicals/Jiangsu Key Laboratory for Food Quality and Safety,Nanjing 210014,China;Guotai Center of Technology Innovation for Veterinary Biologicals,Taizhou 225300,China)

机构地区：[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 [2]兽用生物制品国泰技术创新中心,江苏泰州225300

出　　处：《江苏农业学报》2024年第1期141-148,共8页Jiangsu Journal of Agricultural Sciences

基　　金：江苏省农业科技自主创新基金项目[CX(21)3135];十四五重点研发专项(2022YFD1800800)。

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7 d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。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-carboxymethyl chitosan(OCS)and dextran sulfate(DS),which also had adjuvant activity,were used as carriers to form nanoparticles 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 preparation,and the protein encapsulation rate was 86.56%.The simulated release tests in vitro showed that CTA1-DD protein 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 antibodies 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 

分 类 号：S852.4[农业科学—基础兽医学]