摘要
采用双水相法纯化抗克伦特罗单克隆抗体(CLMcAb),并以其与酶标二抗的亲和力为指标,对CLMcAb的固定化工艺进行研究。在此基础上,通过耐冲刷实验及原子力显微镜(AFM)来表征固定化CLMcAb的稳定性。结果表明:当固定时间90min,CLMcAb稀释倍数2000倍,料液比0.65mL/颗,固定温度4℃时,固定化CLMcAb所表现出的活力最高,即OD值为1.43。纯化介质聚乙二醇(PEG)可通过在载体表面形成均匀紧密的网条结构,并降低其表面粗糙度(Ra)来提高固定化CLMcAb的稳定性。
The anti- clenbuterol monoclonal antibody （CLMcAb） was purified by the aqueous two- phase systems （ATPS） ,and its affinity with the second enzyme-labeled antibody was taken as evaluation indexes to optimize the immobilized process for CLMcAb. Furthermore, the stability of immobilized CLMcAb could be characterized by resist erode test and atomic force microscope （AFM）.The results suggested that the optimum CLMcAb activity could be obtained with the fixed time of 90min,the 2000 fold dilution of CLMcAb and the ratio of solid to liquid of 0.65mL per carrier at 4~C.The immobilization of CLMcAb in PEG phase resulted in the highest activity and stability by reducing the surface roughness（Ra） significantly and forming network micelle on the carrier surface.
出处
《食品工业科技》
CAS
CSCD
北大核心
2014年第7期144-148,153,共6页
Science and Technology of Food Industry
基金
国家自然科学基金(31101283)资助
关键词
CLMcAb
固定化
纯化
响应面
条件优化
anti-clenbuterol monocIonal antibody
immobilization
purification
response surface methodology
conditions optimization
