抗日本血吸虫SIEA26-28KU-scFv-吡喹酮长循环免疫脂质体的制备

发布时间：2018-04-12 15:35

本文选题：日本血吸虫 + 免疫脂质体　；参考：《中南大学》2011年硕士论文

【摘要】：目的制备针对日本血吸虫的第三代吡喹酮长循环免疫脂质体,为日本血吸虫病的靶向性治疗奠定工作基础。方法 1、大量表达并纯化SIEA26-28 ku-scFv 大量培养带有pET32a/SIEA26-28 ku-scFv质粒的BL21(DE3)大肠杆菌,IPTG诱导SIEA26-28 ku-scFv表达,用Ni2+柱纯化目的蛋白,将纯化蛋白进行蛋白浓度测定、SDS-PAGE检测、Western Blot鉴定。 2、制备吡喹酮长循环脂质体采用薄膜蒸发超声分散法制备吡喹酮长循环脂质体并测定其包封率。 3、制备携带单链抗体的长循环免疫脂质体采用新型异型双功能交联剂SPDP交联抗体与脂质体,制备第三代包裹吡喹酮的长循环免疫脂质体,计算抗体偶联率。结果 1、大量表达SIEA26-28 ku-scFv,通过Ni2+螯合亲和层析法提纯单链抗体,通过Western Blot检测证明单链抗体能特异性识别日本血吸虫SIEA。 2、成功制备吡喹酮长循环脂质体,计算其包封率为56.1%。 3、成功制备携带单链抗体与吡喹酮的第三代长循环免疫脂质体,计算抗体偶连率为21.3%。结论成功提取和纯化单链抗体,制备携带单链抗体和吡喹酮的第三代长循环免疫脂质体,为下一步行靶向治疗血吸虫病奠定了工作基础。
[Abstract]:PurposeTo prepare the third generation praziquantel long circulating immune liposomes for schistosomiasis japonicum and lay a foundation for the targeted treatment of schistosomiasis japonicum.Method1. Express and purify SIEA26-28 ku-scFv in large quantitiesThe expression of SIEA26-28 ku-scFv was induced by E. coli BL21 (DE3) with pET32a/SIEA26-28 ku-scFv plasmid. The purified protein was purified by Ni2 column. The protein concentration was determined by SDS-PAGE and identified by Western Blot.Preparation of long circulating liposomes of praziquantelThe long circulating liposomes of praziquantel were prepared by evaporation ultrasonic dispersion method and the encapsulation efficiency was measured.3. Preparation of long circulating immune liposomes carrying scFvThe third generation long circulating immune liposomes coated with praziquantel were prepared by using a new type of bifunctional crosslinking agent SPDP crosslinking antibody with liposome. The coupling rate of antibody was calculated.Result1. SIEA26-28 ku-scFv was expressed in large quantities and purified by Ni2 chelating affinity chromatography. The detection of Western Blot showed that scFv could recognize Schistosoma japonicum SIEA specifically.2. The long circulating liposomes of praziquantel were successfully prepared and the encapsulation efficiency was calculated to be 56.1.3. The third generation of long-cycle immune liposomes carrying single-chain antibody and praziquantel were successfully prepared, and the antibody coupling rate was calculated to be 21.3%.ConclusionThe third generation of long-circulating immune liposomes carrying single-chain antibodies and praziquantel were successfully extracted and purified, which laid a foundation for the further targeted treatment of schistosomiasis.
【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R392

【引证文献】