摘要
目的：制备用于特异性分离阪崎肠杆菌的免疫磁性壳聚糖微球以及对阪崎肠杆菌的捕获效果。方法：采用反向悬浮交联法制备磁性壳聚糖微球．并对微球的表面进行氨基化修饰。然后与阪崎肠杆菌多克隆抗体进行偶联，制备免疫磁性壳聚糖微球。优化磁性壳聚糖微球对阪崎肠杆菌多克隆抗体的偶联条件，包括偶联时间、磁性壳聚糖微球用量、偶联体系pH，对抗体的饱和偶联量。通过与显色培养基结合的方法研究免疫磁性壳聚糖微球对阪崎肠杆菌的捕获能力。结果：制备的磁性壳聚糖微球通过表面修饰能连接上大量的活性氨基。在pH7．4的偶联体系中，10．0min即可完成对阪崎肠杆菌多克隆抗体的偶联，0．01g磁性壳聚糖微球对抗体的饱和偶联量为25～50μL。当阪崎肠杆菌浓度较低时，制备的免疫磁性壳聚糖微球对阪崎肠杆菌的捕获率达94．7％，对阪崎肠杆菌的灵敏度为5cfu／mL。结论：获得阪崎肠杆菌免疫磁性壳聚糖微球，该微球是一种非常有潜力的快速富集、分离阪崎肠杆菌的有效方法。
Objective： The immunomagnetic chitosan beads which separated Enterobacter sakazakii specifically were prepared and the capture capacity of immunomagnetic chitosan beads was studied. Methods： The magnetic chitosan beads were prepared by using reverse phase suspension cross-linking polymerization. Amino modification was done on the sur- face of the magnetic chitosan bead. Then ethanediamin-modified magnetic chitosan beads coupled with polyclonal antibody of Enterobacter sakazakii to get immunomagnetic chitosan beads. The optimal conditions of coupling reaction for ethanedi- amin-modified magnetic chitosan bead and polyclonal antibody were studied including coupling time, the amount of ethanediamin-modified magnetic chitosan beads, coupling pH, the amount of the antibody. The capture capacity of im- munomagnetic chitosan beads were studied with combining to the color medium. Results： The surface of the magnetic chitosan beads had been successfully modified with active ethanediamine. With pH of the coupling buffer of 7.4, poly- clonal antibody of Enterobacter sakazakii was coupled with 0.01 g ethanediamin-modified chitosan magnetic beads for 10.0 min and the amount of coupled antibody was 25uL-50 uL. So the immunomagnetic chitosan beads was got. When in the low contention of the Enterobacter sakazakii the capture ratio of immunomagnetic chitosan beads was about 94.7%, the sensitivity was about 5 cfu/mL. Conclusion： The immunomagnetic cbitosan beads for Enterobacter sakazakii were pre- pared successfully and the bead was a potential method to enrich and separate Enterobacter sakazakii fast.
出处
《中国食品学报》
EI
CAS
CSCD
北大核心
2016年第9期95-100,共6页
Journal of Chinese Institute Of Food Science and Technology
基金
高等学校博士学科点专项科研基金(20110132110007)
国家自然科学基金项目(31371731)
教育部长江学者创新团队(IRT1188)
关键词
壳聚糖
免疫磁性微球
阪崎肠杆菌
分离
chitosan
immunomagnetic beads
Enterobacter sakazakii
separate
