摘要
目的以肝细胞生长因子为包被对象,研究胰岛新生相关蛋白(INGAP)空间稳定免疫脂质体的制备方法、理化性质和体外靶向性。方法制备含PDP-PEG-HSPE的空间稳定脂质体,共价连接马来酰亚胺衍生化IN-GAP。激光散射粒度仪测定其粒径分布,比色法测定包封率,透析法检测药物释放度及累积泄漏率,ELISA法检测脂质体表面的抗体免疫活性,流式细胞仪及免疫荧光考察该免疫脂质体对ARIP细胞(大鼠胰腺导管癌细胞系)的结合活性。结果INGAP-SIL[HGF]脂质体的粒径分布为(95.86±6.54)nm;包封率为(88.37±3.58)%;24h释放度为(23.26±2.45)%;4℃保存时7d内药物累积泄漏率小于3%;4℃和25℃条件下保存7d,免疫活性均在75%以上;INGAP连接到脂质体表面后,其免疫活性基本保留;空间稳定脂质体能特异性地结合ARIP细胞。结论通过PDP-PEG-HSPE共价连接INGAP制备的INGAP-SIL理化性质较稳定,能基本保留INGAP的免疫活性,具有体外靶向细胞抗原的能力。
Objective To study the preparation methods,physical and chemical properties of the islet neogenesis-associated protein （INGAP） sterically stabilized immunoliposomes. Methods An endgroup functionalized polyethylene glycol-lipid deirvative （pyridylthiopropionoylamino-polyethylene glycol-hydrogenated soy phosphatidylethanolamine,PDP-PEG-HSPE） was synthesized and incorporated to sterically stabilized liposomes. After mild thiolysis of the PDP groups by dithiothreitol,liposomes were covalently linked with maleimide-derivatized INGAP and formed sterically stabilized immunoliposomes. The size dis- tribution, immunoreactivity of INGAP-modified sterically stabilized liposomes （INGAP-SIL[ HGFI ） and specific binding properties of the INGAP-SIL[ HGF] to ARIP cells were determined, separately. Results The INGAP-SIL[ HGF] had a narrow size distribution after extrusion and the mean size of this immu- noliposomes was （95.86 ＋ 6.54） nm, encapsulation efficiency was （ 88.37 ＋ 3.58） %. The immunoreac- tivity of INGAP can be preserved after efficient attachment of maleimide-derivatized INGAP to the surface of liposomes. But calculated per antibody concentration, the immunoreactivity of INGAPP-SIL [ HGF ] would obviously decrease compared to that of INGAP-SIL[ HGF]. Significantly higher binding of INGAp-SIL to fixed ARIP cells directed by INGAP was obtained compared to other preparations. Conclusion properties INGAP-SIL [ HGF] prepared by PDP-PEG-HSPE method maintains stabile physicolchemical and most immunoreactivity of INGAP and is able to bind nuclear antigens in vitro.
出处
《哈尔滨医科大学学报》
CAS
北大核心
2009年第6期527-531,共5页
Journal of Harbin Medical University
基金
国家自然科学基金资助项目(30671000)
关键词
胰岛新生相关蛋白
空间稳定免疫脂质体
免疫活性
细胞靶向性
islet neogenesis-associated protein
sterically stabilized immunoliposome
immunoreactivity
targeting
