Department of Chemistry, Sapienza Università di Roma, Box n° 34-Roma 62, Piazzale Aldo Moro 5, 00185 Rome, Italy.
Anal Bioanal Chem. 2010 Dec;398(7-8):2895-903. doi: 10.1007/s00216-010-4104-y. Epub 2010 Sep 22.
Plasma protein adsorption is regarded as a key factor in the in vivo organ distribution of intravenously administered drug carriers, and strongly depends on vector surface characteristics. The present study aimed to characterize the "protein corona" absorbed onto DC-Chol-DOPE cationic liposomes. This system was chosen because it is one of the most efficient and widely used non-viral formulations in vitro and a potential candidate for in vivo transfection of genetic material. After incubation of human plasma with cationic liposomes, nanoparticle-protein complex was separated from plasma by centrifugation. An integrated approach based on protein separation by one-dimensional 12% polyacrylamide gel electrophoresis followed by the automated HPLC-Chip technology coupled to a high-resolution mass spectrometer was employed for protein corona characterization. Thirty gel lanes, approximately 2 mm, were cut, digested and analyzed by HPLC-MS/MS. Fifty-eight human plasma proteins adsorbed onto DC-Chol-DOPE cationic liposomes were identified. The knowledge of the interactions of proteins with liposomes can be exploited for future controlled design of colloidal drug carriers and possibly in the controlled creation of biocompatible surfaces of other devices that come into contact with proteins in body fluids.
血浆蛋白吸附被认为是静脉给予药物载体在体内器官分布的关键因素，并且强烈依赖于载体表面特性。本研究旨在表征吸附在 DC-Chol-DOPE 阳离子脂质体上的“蛋白质冠”。选择该系统是因为它是体外最有效和最广泛使用的非病毒制剂之一，并且是体内转染遗传物质的潜在候选者。在人血浆与阳离子脂质体孵育后，通过离心将纳米粒子-蛋白质复合物从血浆中分离出来。采用基于一维 12%聚丙烯酰胺凝胶电泳的蛋白质分离的综合方法，随后采用自动 HPLC-Chip 技术与高分辨率质谱仪相结合，用于蛋白质冠的表征。分离出 30 个凝胶条带，约 2 毫米宽，通过 HPLC-MS/MS 进行切割、消化和分析。鉴定出 58 种吸附在 DC-Chol-DOPE 阳离子脂质体上的人血浆蛋白。对蛋白质与脂质体相互作用的了解可用于未来胶体药物载体的受控设计，并可能用于控制与体液中蛋白质接触的其他设备的生物相容性表面的形成。
