Differential analysis of "protein corona" profile adsorbed onto different nonviral gene delivery systems
Differential analysis of "protein corona" profile adsorbed onto different nonviral gene delivery systems
复制标题
DOI：
10.1016/j.ab.2011.08.003
复制
发表时间：
2011-12-15
影响因子：
2.9
通讯作者：
Lagana, Aldo
中科院分区：
文献类型：
--
作者：
Capriotti, Anna Laura;Caracciolo, Giulio;Lagana, Aldo
鸟枪蛋白质组学方法被用来表征和比较的蛋白质，导致形成一个丰富的“蛋白质冠”吸附到表面的阳离子脂质体（CL），lipoplexes，脂质/聚阳离子/DNA（LPD）复合物，当他们接触血浆。从血浆中分离纳米颗粒-蛋白质复合物后，消化蛋白质混合物，并通过纳米液体色谱-Orbitrap LTQ-XL质谱法分析肽。与脂复合物结合的蛋白质数量是CL冠中鉴定的蛋白质数量的两倍（208对105），而77种蛋白质是两种冠共同的。结合到LPD复合物表面的蛋白质的数量（其中158、133个是lipoplexes共有的）介于CL和lipoplexes的蛋白质冠中发现的蛋白质之间。其中约一半存在于CLs的蛋白质冠中。通过重叠三种制剂，可以看出，只有12种蛋白质是LPD复合物特有的。这些结果可能有助于设计能够结合最小可能量的蛋白质的基因递送系统，这些蛋白质对转染产生负面影响，选择性地结合能够帮助在体内将载体导向靶点的蛋白质，并获得更有效和更有效的基因治疗。(C)2011 Elsevier Inc. All rights reserved.
A shotgun proteomics approach was used to characterize and compare the proteins that lead to the formation of a rich "protein corona" adsorbed onto the surfaces of cationic liposomes (CLs), lipoplexes, and lipid/polycation/DNA (LPD) complexes, when they come into contact with plasma. After separation of the nanoparticle-protein complex from plasma, the protein mixture was digested, and peptides were analyzed by nanoliquid chromatography-Orbitrap LTQ-XL mass spectrometry. The number of proteins bound to lipoplexes was double that of those identified in the corona of CLs (208 vs 105), while 77 proteins were common to both coronas. The number of proteins bound to the surface of the LPD complexes (158, 133 of which are common to lipoplexes) is intermediate between those found in the protein corona of both CLs and lipoplexes. About half of them were found in the protein corona of CLs. By overlapping the three formulations, it can be seen that only 12 proteins are peculiar to LPD complexes. These results may help in designing gene delivery systems capable of binding the minimum possible quantity of proteins that influence transfection negatively, binding selectively proteins capable of helping in steering in vivo the vector toward the target, and obtaining more efficient and effective gene therapy. (C) 2011 Elsevier Inc. All rights reserved.