海藻酸钠-壳聚糖P-VP8~*IgY微胶囊的制备及其释放性能

Preparation of P-VP8~* IgY-loaded sodium alginatechitosan microcapsules and evaluation of its release performance

目的在前期试验成功制备纯化出抗诺如和轮状病毒的双价特异性P-VP8~*IgY基础上,本研究作者制备海藻酸钠-壳聚糖P-VP8~*IgY微胶囊并优化最佳制备工艺。方法以海藻酸钠-壳聚糖为微胶囊囊材,采用复凝聚技术"一步法"制备P-VP8~*IgY微胶囊。以微胶囊的成囊效果和包封率为评价指标,通过单因素分析和L9(34)正交试验优化微胶囊的最佳制备工艺。并运用SDS-PAGE和ELISA法评价海藻酸钠-壳聚糖P-VP8~*IgY微胶囊的释放性能和生物学活性。结果海藻酸钠-壳聚糖P-VP8~*IgY微胶囊的最佳制备工艺条件为:海藻酸钠浓度为3. 0%、壳聚糖浓度为0. 4%、氯化钙浓度为2%(均为质量分数)、壳聚糖与氯化钙溶液(成囊液)的p H值为5. 5,IgY投药量为10 g·L-1。制备的海藻酸钠-壳聚糖P-VP8~*IgY微胶囊大部分呈规则球形,表面较为光滑圆整,包封率可达到85%。制备的微胶囊在胃液2 h后的释放率为7%,肠液6 h后的释放率高达86%,且生物活性良好。结论本工艺制备的海藻酸钠-壳聚糖P-VP8~*IgY微胶囊能够保护特异性P-VP8~*IgY不被胃酸环境破坏,具有较好的缓释作用。

Objective On the basis of successful preparation of purified P-VP8-＊dual IgY specific to norovirus and rotavirus,this study aimed to establish the optimal preparation process of microcapsules and evaluate its sustained release performance. Methods P-VP8-＊IgY microcapsules were prepared by one-step method using chitosan and sodium alginate as wall materials. Preparation of the microcapsules was evaluated by cystoid shape together withencapsulation rate and optimized using single factor analysis and L9（ 34）orthogonal design. The release rate and bio-activity of IgY were also evaluated using SDS-PAGE and ELISA. Results The concentration optimal conditions were showed as follows： 3. 0% sodium alginate,0. 3%chitosan,2. 0% calcium chloride（ mass fraction）,the pH value of chitosan and calcium chloride solution5. 5,and IgY loading rate 10 g·L^-1. M ostof microcapsules shaped round and smooth,the encapsulationratec could reach to 85%. The release rate was only 7. 1% after 2 h simulated gastric fluid digestion,while it could reach more than 86% after 6 h simulated intestinal fluid digestion. Conclusion The chitosan-alginate P-VP8-＊IgY microcapsules had a good sustained release effect,which can protect P-VP8-＊IgY from destruction in gastric acid,and achieve the sustained release in intestinal tract.