Abstract
Immunoglobulin Y (IgY) is an effective orally administered antibody used to protect against various intestinal pathogens, but which cannot tolerate the acidic gastric environment. In this study, IgY was microencapsulated by alginate (ALG) and coated with chitooligosaccharide (COS). A response surface methodology was used to optimize the formulation, and a simulated gastrointestinal (GI) digestion (SGID) system to evaluate the controlled release of microencapsulated IgY. The microcapsule formulation was optimized as an ALG concentration of 1.56% (15.6 g/L), COS level of 0.61% (6.1 g/L), and IgY/ALG ratio of 62.44% (mass ratio). The microcapsules prepared following this formulation had an encapsulation efficiency of 65.19%, a loading capacity of 33.75%, and an average particle size of 588.75 µm. Under this optimum formulation, the coating of COS provided a less porous and more continuous microstructure by filling the cracks on the surface, and thus the GI release rate of encapsulated IgY was significantly reduced. The release of encapsulated IgY during simulated gastric and intestinal digestion well fitted the zero-order and first-order kinetics functions, respectively. The microcapsule also allowed the IgY to retain 84.37% immune-activity after 4 h simulated GI digestion, significantly higher than that for unprotected IgY (5.33%). This approach could provide an efficient way to preserve IgY and improve its performance in the GI tract.
摘 要
目 的
制备、 优化、 表征和体外评价包埋免疫球蛋白 Y (IgY) 的壳寡糖-海藻酸盐微胶囊, 以实现 IgY 的胃肠道控释, 提高 IgY 胃肠道消化过程中的活性保持率.
创新点
以壳寡糖为涂层, 采用两步法制备包埋 IgY 的微胶囊; 采用响应面法全面优化了微胶囊的配方工艺; 对 IgY 的体外控释进行了系统的评价.
方 法
通过两步离子凝胶法制备包埋 IgY 的微胶囊, 采用响应面法优化微胶囊配方工艺, 通过包埋率、 负载率、 平均粒径、 微观形貌、 色度和球形度等指标表征微胶囊, 采用模拟胃肠道消化体系体外评价 IgY 的释放动力学和免疫活性保持率.
结 论
采用响应面法优化的微胶囊配方工艺为海藻酸钠浓度1.56%(15.6 g/L)、 壳寡糖浓度 0.61%(6.1 g/L) 和 IgY/海藻酸钠比率 62.44% (质量比), 由此制备的微胶囊包埋率达 65.19%, 负载率达 33.75%, 平均粒径为 588.75 μm. 壳寡糖涂层通过填充效应和静电相互作用使微胶囊表面更加光滑和连续, 从而显著降低了模拟胃肠道消化过程中 IgY 的释放速率. 微胶囊中 IgY 的模拟胃部控释和模拟肠道控释分别符合零级动力学方程和一级动力学方程 (R2 > 0.99). 微胶囊在模拟胃肠道消化 4 h 后的 IgY 免疫活性保持率达 84.37%, 远高于未进行微胶囊化保护的 IgY (5.33%).
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Acknowledgments
The authors thank Mrs. Chun-jiao SONG and Mrs. Zhenfeng LIAO (Electronic Imaging Laboratory, Public Experimental Platform, Zhejiang Academy of Agricultural Sciences, Hangzhou, Zhejiang, China) for their advice and support on the surface morphology observation of microcapsules.
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Jin ZHANG performed the experimental research and data analysis, wrote and edited the manuscript. Huan-huan LI, Yi-fan CHEN, Yun-xin YAO, and Xu-ming LIU performed the data analysis. Qian LAN and Xiao-fan YU collected and analyzed the data. Li-hong CHEN and Hong-gang TANG contributed to the study design and data analysis. Fan-bin KONG contributed to the writing and editing of the manuscript. All authors have read and approved the final manuscript and, therefore, have full access to all the data in the study and take responsibility for the integrity and security of the data.
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Jin ZHANG, Huan-huan LI, Yi-fan CHEN, Li-hong CHEN, Hong-gang TANG, Fan-bin KONG, Yun-xin YAO, Xu-ming LIU, Qian LAN, and Xiao-fan YU declare that they have no conflict of interest.
No studies using human or animal subjects were performed by any of the authors.
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Project supported by the National Key Research and Development Program of China (No. 2018YFD0400305), the Modern Agro-industry Technology Research System of China (No. CARS-40-K26), and the “One Belt and One Road” International Science and Technology Cooperation Program of Zhejiang, China (No. 2019C04022)
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Zhang, J., Li, Hh., Chen, Yf. et al. Microencapsulation of immunoglobulin Y: optimization with response surface morphology and controlled release during simulated gastrointestinal digestion. J. Zhejiang Univ. Sci. B 21, 611–627 (2020). https://doi.org/10.1631/jzus.B2000172
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DOI: https://doi.org/10.1631/jzus.B2000172
Key words
- Immunoglobulin Y (IgY)
- Microencapsulation
- Chitooligosaccharide (COS)
- Response surface methodology (RSM)
- Controlled release
- Simulated gastrointestinal digestion (SGID)