穿膜肽-抗体-微球偶联物的制备方法研究  

Studies on Preparation Method of CCPs-Antibody-Nanospheres Conjugates

作　　者：王启辉[1] 刘金华[1] 周洁[2] 冷静[3] 

机构地区：[1]广西医科大学免疫学教研室 [2]广西医科大学第一附属医院泌尿外科,南宁530021 [3]广西中医学院,南宁530001

出　　处：《黑龙江医药》2011年第4期558-561,共4页Heilongjiang Medicine journal

基　　金：国家自然科学基金(30700836)

摘　　要：目的:探讨制备穿膜肽-抗体-微球新型药物传送系统的可行性。方法:采用N-琥珀酰亚胺基-3-(2-吡啶二硫)-丙酸酯(SPDP)交联法将穿膜肽-增强型绿色荧光蛋白融合蛋白(CPPs-EGFP)、牛血清白蛋白微球(BSA-NS)、乙肝高效价免疫球蛋白(HBIg)分别采用一次偶联法和二次偶联法两种交联方案进行共价交联,采用还原和非还原SDS聚丙烯酰胺凝胶电泳(SDS-PAGE)和免疫荧光法评价穿膜肽、抗体、微球间的交联。结果:两种交联方案制备的穿膜肽-抗体-微球偶联物,SDS-PAGE还原电泳均可见在低分子量端形成三条蛋白条带,非还原电泳均未见条带;在不同激发波长的荧光显微镜下,均可见微球表面分别发出绿色荧光和红色荧光。结论:应用SP-DP交联剂,无论是一次偶联法还是二次偶联法均能够将穿膜肽、抗体、微球有效地进行偶联,为进一步研究穿膜肽-抗体-微球新型药物传送系统的特性奠定了基础。Objective：To explore the feasibility of applying N-succinimidyl-3-（2-pyridyldithio） propionate（SPDP） conjugation technology to the preparation of a new drug delivery system：CCPs-Antibody-Nanospheres conjugates.Methods：CCPs-Antibody-Nanospheres conjugates were prepared by two different methods of SPDP cross-linking. CCPs-Antibody-Nanospheres conjugates were tested by SDS-PAGE assay and immunofluorescent assay.Results：CCPs-Antibody-Nanospheres conjugates prepared by two different methods,which were reduced with DTT all showed three protein bands at low molecular weight end. There was brightest reaction in immunofluorescent.Conclusion：These data implicated that conjunction of cell-penetrating peptides,antibodies and nanospheres by two different methods of SPDP conjugation technology was successful and this would provide a basis for further studies on the new drug delivery system：CCPs-Antibody-Nanospheres conjugates.

关 键 词：抗体 穿膜肽 牛血清白蛋白 微球 蛋白偶联 SPDP 

分 类 号：R392[医药卫生—免疫学]