Northwestern University, Feinberg School of Medicine, Department of Urology, Chicago, Illinois 60611, United States.
Northwestern University, Simpson Querrey Institute for Bionanotechnology (SQI), Chicago, Illinois 60611, United States.
ACS Sens. 2020 Oct 23;5(10):3019-3024. doi: 10.1021/acssensors.0c01117. Epub 2020 Jul 14.
High-density lipoproteins (HDL) are constitutionally dynamic nanoparticles that circulate in the blood. The biological functions of HDLs are impacted by interchangeable surface chemical components, like cholesterol and HDL-associated proteins. Current methods to quantify the chemical constituents of HDL are largely restricted to clinical or academic laboratories and require expensive instrumentation, and there is no commonality to the techniques required to detect and quantify different analytes (e.g., cholesterol versus HDL-associated protein). To potentially facilitate and streamline the analysis of HDL composition, we hypothesized that mixing native HDLs with similarly sized gold nanoparticles whose surfaces are endowed with phospholipids, called complementary nanoparticle scaffolds (CNS), would enable interparticle exchange of surface components. Then, easy isolation of the newly formed particles could be accomplished using benchtop centrifugation for subsequent measurement of HDL components exchanged to the surface of the CNS. As proof-of-concept, data demonstrate that CNS incubated with only a few microliters of human serum rapidly (1 h) sequester cholesterol and HDL-associated proteins with direct correlation to native HDLs. As such, data show that the CNS assay is a single platform for rapid isolation and subsequent detection of the surface components of native HDLs.
高密度脂蛋白 (HDL) 是构成上动态的纳米颗粒,在血液中循环。HDL 的生物学功能受可互换的表面化学组成部分(如胆固醇和 HDL 相关蛋白)的影响。目前定量测定 HDL 化学成分的方法在很大程度上仅限于临床或学术实验室,并且需要昂贵的仪器,并且检测和定量不同分析物(例如胆固醇与 HDL 相关蛋白)所需的技术没有共性。为了可能促进和简化 HDL 组成的分析,我们假设将天然 HDL 与表面赋予磷脂的类似大小的金纳米颗粒(称为互补纳米颗粒支架 (CNS))混合,将能够实现颗粒间表面成分的交换。然后,可以使用台式离心机轻松分离新形成的颗粒,随后测量 CNS 表面交换的 HDL 成分。作为概念验证,数据表明,仅用几微升人血清孵育的 CNS 迅速(1 小时)隔离胆固醇和 HDL 相关蛋白,与天然 HDL 直接相关。因此,数据表明 CNS 测定法是用于快速分离和随后检测天然 HDL 表面成分的单一平台。