ACPP-pHPMA-Ad.egfp接合物的合成及其逃逸腺病毒中和抗体的作用检测  

Construction of ACPP-pHPMA-Ad.egfp conjugate and study on its ability to evade neutralising antibody

作　　者：李书华[1] 陈婷婷[1] 刘谕昆 李灏[1] 陈贤亮[1] 张雅洁[1] 

机构地区：[1]广州医科大学,广东广州510182

出　　处：《中国医院药学杂志》2014年第11期859-863,共5页Chinese Journal of Hospital Pharmacy

基　　金：广东省医学科研基金项目(编号:A2013244)

摘　　要：目的:合成一种由可活化细胞穿膜肽ACPP、水溶性高分子聚合物pHPMA修饰腺病毒的接合物,并初步探索该接合物逃逸腺病毒中和抗体免疫清除的能力。方法:通过化学合成的方法合成高分子聚合物pHPMA及可活化细胞穿膜肽ACPP,通过化学合成的方法合成了接合物ACPP-pc-Ad.egfp。通过动态光散射检测高分子聚合物ACPP-pc-Ad.egfp的粒径,通过酶标仪检测接合物ACPP-pc-Ad.egfp感染能力,通过酶标仪检测接合物ACPP-pc-Ad.egfp感染能力免疫逃逸能力。结果:成功合成了高分子聚合物pHPMA及可活化细胞穿膜肽ACPP;动态光散射检测结果表明ACPP及pHPMA修饰的腺病毒接合物粒径增大;酶标仪检测接合物ACPP-pc-Ad.egfp具有较强的感染A549细胞的能力,并对腺病毒中和抗体的免疫清除发生了明显的免疫逃逸。结论:成功合成了ACPP-pc-Ad.egfp接合物,并证实ACPP-pc-Ad.egfp能对机体的腺病毒免疫清除发生免疫逃逸。OBJECTlVE To synthesize and characterize an ACPP and multivalent hydrophilic HPMA polymer-coated adeno- virus conjugate, for increase the resistance of adenovirus to antibody neutralization. METttODS A random poly-[-N-（2- hydroxypropyl） methacrylamide~ （pHPMA） and activable cell-penetrating peptide （ACPP） were prepared by free radical pre- cipitation copolymerization method, polymer modification of virus was performed by incubating ACPP,pHPMA and adenovirus in 10% glycerol/PBS （pH 7. 8） at 4 ℃ for 12 h to form ACPP-pc- Ad. egfp conjugates Particle size after virus modification was measured by dynamic light scattering （DLS） the infected ability of ACPP-pc- Ad. egfp was measured at λex 488 nm and 2,m 538 nm in a Fluoroskan plate reader, and virus neutralisation assay of ACPP -pc-Ad. egfp was examined by measuring gfp ex- pression in A549 cells. RESULTS pHPMA and peptides ACPP were synthesized as devised and ACPP -pc-Ad. egfp was pre pared successfully, the average diameter of ACPP-pc- Ad. egfp was bigger than that of Ad. egfp and pc Ad. egfp demonstrated that efficient virus modification have been achieved ; gfp expression measured in a fluorescence plate reader indicated that ACPP- pc-Ad, egfp conjugates kept the ability to infect A549 cells and virus neutralisation assays showed that ACPP-pc-Ad. egfp was 9-fold more resistant to neutralisation than Ad. egfpo CONCLUSION Polymer-coated adenovirus ACPP-pc-Ad. egfp was suc- cessfully synthesized, the virus neutralisation assay Results showed ACPP-pc-Ad. egfp could evade neutralising antibody effi- ciently.

关 键 词：pHPMA ACPP ACPP-pc—Ad EGFP 免疫逃逸 

分 类 号：R963[医药卫生—微生物与生化药学]