摘要
采用合成的表面氨基化磁性纳米颗粒,通过化学共价交联制备了葡萄球菌ProteinA磁性纳米颗粒载体(SPA-MP),探讨了载体制备的优化条件,根据生物分子特异性亲合作用原理,在外加磁场的定向控制下,通过亲和吸附、清洗和解吸附等操作,探讨了SPA-MP载体在抗体分离纯化领域的应用可行性。载体制备优化实验结果显示,通过改变蛋白质浓度、交联剂浓度和交联剂活化时间可以制备不同表面密度的SPA-MP载体。300μgSPA,2.5%(V/V)戊二醛浓度和3h的活化时间可以获取表面密度高达35mgSPA/g磁性纳米颗粒的载体。此外,应用结果显示每克SPA-MP磁性微球载体可以结合约15～18mg的CD25抗体,同时可有效地分离纯化人抗血清样品中的IgG抗体。
Surface amino-modified magnetic nanosparticle was prepared. Staphylococcal protein A （SPA） coated magnetic nanosphere carrier （SPA-MP） was prepared by covalent coupling method, and the optimization condition for the preparation of carrier was discussed. Then according to the specific affinity between biological molecules, application feasibility of the SPA-MP carrier for antibody purification field was investigated through the oriented controlling of additional magnetic field, affinity adsorption, washing and elution operations. The optimization experiments appeared that SPA-MP carriers with different surface density can be prepared by changing the protein concentration, cross -linker concentration and activation time of cross -linker. Under the conditions of 300μg SPA ,2.5% （V/V） glutaraldehyde concentration and 3h activation time, the SPA-MP cartier with surface density as high as 35 mg SPA per gram magnetic nanosphere can be prepared. In addition, application results indicated that 1 g SPA-MP cartier can bind about 15 -18mg CD25 monoclonal antibody, and purified IgG antibody from human antiserum samples effectively.
出处
《中国生物工程杂志》
CAS
CSCD
北大核心
2008年第1期13-17,共5页
China Biotechnology
