NanoSight Limited Release: Method Development to Estimate the Purity of Vesicle Preparations Using Nanoparticle Tracking Analysis at the Cardiff University
Published: Feb 26, 2013
The School of Medicine at Cardiff University is one of the largest medical schools in the United Kingdom, home to over 3000 students and staff. It is a major international center for teaching and research providing a vibrant community of medical endeavor. The School is based at the University Hospital of Wales in Cardiff, but is also embedded at other hospital sites in Cardiff including Velindre Cancer Centre.
Dr Aled Clayton and his team are researching the roles of exosomes in modulating tumor immunity and the cancer microenvironment in prostate cancer. Exosomes are nanometer sized vesicles, produced in abundance by malignant cells but much of their physiological functions are incompletely understood.
Among Dr Clayton's interests are exosomes as possible disease markers, and has been using the NanoSight nanoparticle characterization system as a reliable tool to help develop projects in this field through the analysis of urine/serum from patients. He is keen to develop fluorescent methods with this platform in the future.
Describing his work, Dr Clayton says "as well as our own interests in cancer, we also collaborate with local colleagues interested in diverse conditions (Alzheimer's, cardiac disease, renal disease and others) in which vesicles may play a role. NanoSight's nanoparticle tracking analysis system has been a versatile platform aiding all of these studies."
Continuing, he said "prior to using NTA, there were few other options available. We had made use of electron microscopy but, we have come to rely on NTA as it is a very sensitive and versatile platform, well-suited to daily use."
The work of Dr Clayton and his postdoc, Dr Jason Webber, has recently been published in the Journal of Extracellular Vesicles where they posed the question "How pure are your vesicles?" In this paper, they proposed a straightforward method to estimate the purity of vesicle preparations by comparing the ratio of nanovesicle counts to protein concentration, using tools including the NanoSight NTA platform and a colorimetric protein assay such as the BCA-assay. They show this approach is simple enough to apply to every vesicle preparation within a given laboratory, assisting researchers as a routine quality control step. The paper also proposes that the approach may aid in comparing/standardising vesicle purity across diverse studies, and may be of particular importance in evaluating vesicular biomarkers.(1)
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NanoSight delivers the world's most versatile and proven multi-parameter nanoparticle analysis in a single instrument.
NanoSight's "Nanoparticle Tracking Analysis" (NTA) detects and visualizes populations of nanoparticles in liquids down to 10 nm, dependent on material, and measures the size of each particle from direct observations of diffusion. Additionally, NanoSight measures concentration and a fluorescence mode differentiates suitably-labelled particles within complex background suspensions. Zeta potential measurements are similarly particle-specific. It is this particle-by-particle methodology that takes NTA beyond traditional light scattering and other ensemble techniques in providing high-resolution particle size distributions and validates data with information-rich video files of the particles moving under Brownian motion.
This simultaneous multiparameter characterization matches the demands of complex biological systems, hence its wide application in development of drug delivery systems, of viral vaccines, and in nanotoxicology. This real-time data gives insight into the kinetics of protein aggregation and other time-dependent phenomena in a qualitative and quantitative manner. NanoSight has a growing role in biodiagnostics, being proven in detection and speciation of nanovesicles (exosomes) and microvesicles.
NanoSight has installed more than 500 systems worldwide with users including BASF, GlaxoSmithKline, Merck, Novartis, Pfizer, Proctor and Gamble, Roche and Unilever together with the most eminent universities and research institutes. NanoSight's technology is validated by 600+ third party papers citing NanoSight results. NanoSight's leadership position in nanoparticle characterization is consolidated further with publication of an ASTM International standard, ASTM E2834, which describes the NTA methodology for detection and analysis of nanoparticles. For more information, visit www.nanosight.com
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