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期刊信息:
《药学服务与研究》2018年, 第18卷, 第2期, 第87-91,95页
标题:
A33抗体修饰的外泌体给药系统的构建及体外评价
DOI:
10.5428/pcar20180202
作者:
1. 李艳1(1.江西中医药大学药学院 南昌 330004 liyan52633@163.com)
2. 2(2.第二军医大学长海医院药学部 上海 200433 liyan52633@163.com)
3. 高申2(2.第二军医大学长海医院药学部 上海 200433 )
4. 夏清明2(2.第二军医大学长海医院药学部 上海 200433 )
5. 宫春爱2(2.第二军医大学长海医院药学部 上海 200433 )
6. 郭慧玲1(1.江西中医药大学药学院 南昌 330004 ghl6262@126.com)
摘要:
摘要  目的:构建A33抗体修饰的外泌体(exosomes,EXO)包载多柔比星(doxorubicin,DOX)给药系统并进行体外评价。方法:通过化学交联法将A33抗体与羧基化的超顺磁性纳米粒(ultrasmall superparamagnetic iron oxide nanoparticles,USPIO)连接制备探针(A33-USPIO),并利用动态光散射和红外光谱法对连接前后的USPIO进行表征。采用超速离心法分离出人结肠癌细胞株LIM1215细胞来源的EXO包载DOX,并通过透射电镜观察形态。用ELISA法测定EXO和探针结合的最适比例。透析法测定A33抗体修饰的载DOX的EXO给药系统(A33-US-EXO/DOX)体外释放特性。将不同的DOX制剂分别与LIM1215共培养后,用流式细胞仪测定细胞摄取DOX的情况,利用荧光显微镜验证A33-US-EXO/DOX的体外靶向能力。结果:透射电镜下EXO直径约为100 nm,2.5 μg 探针约能与32 μg的EXO结合,结合后平均粒径为 (187.83±6.76) nm,聚合物分散指数(polymer dispersity index,PDI)为0.21。在pH 7.4、6.0、5.0条件下,DOX组、EXO/DOX组和A33-US-EXO/DOX组的48 h累积释药量分别为24.32%、34.07%、62.82%,细胞摄取效率分别为(13.10±1.08)%、(51.53±3.56)%和 (85.11±3.91)%。荧光观察显示,A33-US-EXO/DOX与A33抗原阳性的LIM1215细胞大量结合,而与A33抗原阴性的RAW264.7细胞结合较少。结论:本研究成功构建了A33抗原靶向的EXO给药系统,体外实验表明其具有较高的摄取效率和良好的靶向性。
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若需在您的论文中引用此文,请按以下格式著录参考文献:
中文著录格式 李艳1,2,高申2,夏清明2,宫春爱2,郭慧玲1. A33抗体修饰的外泌体给药系统的构建及体外评价[J]. 药学服务与研究. 2018; 18(2): 87-91,95.
英文著录格式 LI Yan1,2,GAO Shen2,XIA QingMing2,GONG ChunAi2,GUO HuiLing1. Construction and in vitro evaluation of A33 antibody functionalized exosomes drug delivery system[J]. Pharmaceutical Care and Research / yao xue fu wu yu yan jiu. 2018; 18(2): 87-91,95.
参考文献:
1. Koppers-Lalic D,Hogenboom M M,Middeldorp J M,et al.Virus-modified exosomes for targeted RNA delivery; a new approach in nanomedicine[J].Adv Drug Deliv Rev,2013,65(3):348-356.
2. Kahlert C,Kalluri R.Exosomes in tumor microenvironment influence cancer progression and metastasis[J].J Mol Med(Berl),2013,91(4):431-437.
3. Mathivanan S,Lim J W E,Tauro B J,et al.Proteomics analysis of A33 immunoaffinity-purified exosomes released from the human colon tumor cell line LIM1215 reveals a tissue-specific protein signature[J].Mol Cell Proteomics,2010,9(2):197-208.
4. Johnstone C N,Tebbutt N C,Abud H E,et al.Characterization of mouse A33 antigen,a definitive marker for basolateral surfaces of intestinal epithelial cells[J].Am J Physiol Gastrointest Liver Physiol,2000,279(3):G500-G510.
5. Deckert P M,Renner C,Cohen L S,et al.A33scFV-cytosine deaminase:a recombinant protein construct for antibody-directed enzyme-prodrug therapy[J].Br J Cancer,2003,88(6):937-939.
6. Cortez C,Tomaskovic-Crook E,Johnston A P R,et al.Influence of size,surface,cell line,and kinetic properties on the specific binding of A33 antigen-targeted multilayered particles and capsules to colorectal cancer cells[J].ACS Nano,2007,1(2):93-102.
7. Carrasquillo J A,Pandit-Taskar N,O’Donoghue J A,et al.124I-huA33 antibody PET of colorectal cancer[J].J Nucl Med,2011,52(8):1173-1180.
8. Sakamoto J,Oriuchi N,Mochiki E,et al.A phase Ⅰ radioimmunolocalization trial of humanized monoclonal antibody huA33 in patients with gastric carcinoma[J].Cancer Sci,2006,97(11):1248-1254.
9. Théry C,Glayton A,Amigorena S,et al.Isolation and characterization of exosomes from cell culture supernatants and biological fluids[J].Curr Protoc Cell Biol,2006:Chapter 3 Unit 3.22.1-3.22.29.
10. Masyuk A I,Masyuk T V,LaRusso N F.Exosomes in the pathogenesis,diagnostics and therapeutics of liver diseases[J].J Hepatol,2013,59(3):621-625.
11. Raposo G,Stoorvogel W.Extracellular vesicles: exosomes,microvesicles,and friends[J].J Cell Biol,2013,200(4):373-383.
12. Kim M S,Haney M J,ZHAO YuLing,et al.Development of exosome-encapsulated paclitaxel to overcome MDR in cancer cells[J].Nanomedicine,2016,12(3):655-664.
13. QI HongZhao,LIU ChaoYong,LONG LiXia,et al.Blood exosomes endowed with magnetic and targeting properties for cancer therapy[J].ACS Nano,2016,10(3):3323-3333.
14. Toffoli G,Hadla M,Corona G,et al.Exosomal doxorubicin reduces the cardiac toxicity of doxorubicin[J].Nanomedicine(Lond),2015,10(19): 2963-2971.
15. Agrawal A K,Aqil F,Jeyabalan J,et al.Milk-derived exosomes for oral delivery of paclitaxel[J].Nanomedicine,2017,13(5):1627-1636.
16. 许金霞,唐建斌,赵鲁杭,等.肿瘤pH响应的聚合物胶束用于肿瘤药物靶向输送的研究进展[J].药学学报,2009,44(12):1328-1335.