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      Reference materials and representative test materials to develop nanoparticle characterization methods: the NanoChOp project case

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          Abstract

          This paper describes the production and characteristics of the nanoparticle test materials prepared for common use in the collaborative research project NanoChOp (Chemical and optical characterization of nanomaterials in biological systems), in casu suspensions of silica nanoparticles and CdSe/CdS/ZnS quantum dots (QDs). This paper is the first to illustrate how to assess whether nanoparticle test materials meet the requirements of a “reference material” (ISO Guide 30, 2015) or rather those of the recently defined category of “representative test material (RTM)” (ISO/TS 16195, 2013). The NanoChOp test materials were investigated with small-angle X-ray scattering (SAXS), dynamic light scattering (DLS), and centrifugal liquid sedimentation (CLS) to establish whether they complied with the required monomodal particle size distribution. The presence of impurities, aggregates, agglomerates, and viable microorganisms in the suspensions was investigated with DLS, CLS, optical and electron microscopy and via plating on nutrient agar. Suitability of surface functionalization was investigated with attenuated total reflection Fourier transform infrared spectrometry (ATR-FTIR) and via the capacity of the nanoparticles to be fluorescently labeled or to bind antibodies. Between-unit homogeneity and stability were investigated in terms of particle size and zeta potential. This paper shows that only based on the outcome of a detailed characterization process one can raise the status of a test material to RTM or reference material, and how this status depends on its intended use.

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          Most cited references29

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          Mapping protein binding sites on the biomolecular corona of nanoparticles.

          Nanoparticles in a biological milieu are known to form a sufficiently long-lived and well-organized 'corona' of biomolecules to confer a biological identity to the particle. Because this nanoparticle-biomolecule complex interacts with cells and biological barriers, potentially engaging with different biological pathways, it is important to clarify the presentation of functional biomolecular motifs at its interface. Here, we demonstrate that by using antibody-labelled gold nanoparticles, differential centrifugal sedimentation and various imaging techniques it is possible to identify the spatial location of proteins, their functional motifs and their binding sites. We show that for transferrin-coated polystyrene nanoparticles only a minority of adsorbed proteins exhibit functional motifs and the spatial organization appears random, which is consistent, overall, with a stochastic and irreversible adsorption process. Our methods are applicable to a wide array of nanoparticles and can offer a microscopic molecular description of the biological identity of nanoparticles.
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            Common pitfalls in nanotechnology: lessons learned from NCI's Nanotechnology Characterization Laboratory.

            The Nanotechnology Characterization Laboratory's (NCL) unique set-up has allowed our lab to handle and test a variety of nanoparticle platforms intended for the delivery of cancer therapeutics and/or imaging contrast agents. Over the last six years, the NCL has characterized more than 250 different nanomaterials from more than 75 different investigators. These submitted nanomaterials stem from a range of backgrounds and experiences, including government, academia and industry. This has given the NCL a unique and valuable opportunity to observe trends in nanoparticle safety and biocompatibility, as well as note some of the common mistakes and oversights of nanoformulation. While not exhaustive, this article aims to share some of the most common pitfalls observed by the NCL as they relate to nanoparticle synthesis, purification, characterization and analysis.
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              Quantitative profiling of the protein coronas that form around nanoparticles.

              Nanoparticle applications in biotechnology and biomedicine are steadily increasing. In biological fluids, proteins bind to nanoparticles that form the protein corona, crucially affecting the nanoparticles' biological identity. As the corona affects in vitro and/or in vivo nanoparticle applications, we developed a method to obtain time-resolved protein corona profiles formed on various nanoparticles. After incubation in plasma or a similar biofluid, or after injection into a mouse, the first analytical step is sedimentation of the nanoparticle-protein complexes through a sucrose cushion, thereby allowing analysis of early corona formation time points. Next, corona profiles are visualized by gel electrophoresis and quantitatively analyzed after tryptic digestion using label-free liquid chromatography-high-resolution mass spectrometry. In contrast to other approaches, our established methodology allows the researcher to obtain qualitative and quantitative high-resolution corona signatures. The protocol can be readily extended to the investigation of protein coronas from various nanomaterials (as an example, we applied this protocol to different silica nanoparticles (SiNPs) and polystyrene nanoparticles (PSNPs)). Depending on the number of samples, the protocol from nanoparticle-protein complex recovery to data evaluation takes ~8-12 d to complete.
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                Author and article information

                Contributors
                Journal
                Front Chem
                Front Chem
                Front. Chem.
                Frontiers in Chemistry
                Frontiers Media S.A.
                2296-2646
                19 October 2015
                2015
                : 3
                : 56
                Affiliations
                [1] 1Institute for Reference Materials and Measurements, Joint Research Centre, European Commission Geel, Belgium
                [2] 2Institute of Materials and Environmental Chemistry, Research Centre for Natural Sciences, Hungarian Academy of Sciences Budapest, Hungary
                [3] 3Physikalisch-Technische Bundesanstalt Berlin, Germany
                [4] 4LGC Limited Teddington, UK
                [5] 5Biophotonics Division 1.10, Federal Institute for Materials Research and Testing Berlin, Germany
                [6] 6Analytical Sciences, National Physical Laboratory Teddington, UK
                Author notes

                Edited by: Claudia Cascio, RIKILT, Netherlands

                Reviewed by: Jay Nadeau, McGill University, Canada; Isaac Ojea Jimenez, European Comission-Joint Research Centre, Spain

                *Correspondence: Gert Roebben gert.roebben@ 123456ec.europa.eu

                This article was submitted to Analytical Chemistry, a section of the journal Frontiers in Chemistry

                Article
                10.3389/fchem.2015.00056
                4609882
                c8ea1485-e789-43df-bad3-728b793a1590
                Copyright © 2015 Roebben, Kestens, Varga, Charoud-Got, Ramaye, Gollwitzer, Bartczak, Geißler, Noble, Mazoua, Meeus, Corbisier, Palmai, Mihály, Krumrey, Davies, Resch-Genger, Kumarswami, Minelli, Sikora and Goenaga-Infante.

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                History
                : 13 July 2015
                : 11 September 2015
                Page count
                Figures: 5, Tables: 8, Equations: 1, References: 48, Pages: 16, Words: 11630
                Funding
                Funded by: EMRP participating countries of EURAMET and the European Union
                Award ID: EMRP NEW 03
                Categories
                Chemistry
                Technology Report

                nanoparticle,materials characterization,reference material,analytical quality assurance,metrology

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