Center for Photonic Science and Engineering, Skolkovo Institute of Science and Technology, Skolkovo Innovation Center, Building 3, Moscow, 143026, Russian Federation; School of Biological and Medical Physics, Moscow Institute of Physics and Technology, Institutsky per. 9/7, Dolgoprudny, Moscow Region, 141700, Russian Federation.
School of Biological and Medical Physics, Moscow Institute of Physics and Technology, Institutsky per. 9/7, Dolgoprudny, Moscow Region, 141700, Russian Federation.
Biochem Biophys Res Commun. 2022 Jun 18;609:189-194. doi: 10.1016/j.bbrc.2022.04.017. Epub 2022 Apr 10.
The concentration of extracellular vesicles (EVs) is an essential attribute of biofluids and EV preparations. EV concentration in body fluids was correlated with health status. The abundance of EV secreted by cultured cells into growth medium is vital in signaling studies, tissue and disease models, and biomanufacturing of acellular therapeutic secretome. A limited number of physical principles sensitive to EV concertation have been discovered so far. Particle-by-particle counting methods enumerate individual particles scattering light, modulating the Coulter current, or appearing in EM images. The available ensemble techniques in current use rely on the concentration-dependent signal intensity, as in the case of ELISA. In this study, we propose for the first-time the ensemble-based characterization of EV concentration by dynamic surface tension (DST) probe and demonstrate its implementation. We show that DST measurements agree with the widely used NTA measurements of EV concertation. The proposed method is low-cost and requires only basic laboratory equipment for implementation.
细胞外囊泡 (EVs) 的浓度是生物流体和 EV 制剂的一个重要属性。体液中的 EV 浓度与健康状况相关。培养细胞分泌到生长培养基中的 EV 的丰度对于信号研究、组织和疾病模型以及无细胞治疗分泌组的生物制造至关重要。到目前为止,只发现了少数对 EV 浓度敏感的物理原理。逐个粒子计数方法对散射光的单个粒子进行计数,调节库尔特电流,或出现在 EM 图像中。目前使用的现有集合技术依赖于浓度依赖性的信号强度,如在 ELISA 的情况下。在这项研究中,我们首次提出了基于动态表面张力 (DST) 探针的集合特征描述 EV 浓度的方法,并证明了其实施。我们表明,DST 测量结果与广泛使用的 EV 浓度 NTA 测量结果一致。该方法成本低,仅需基本的实验室设备即可实施。