High-throughput profiling of nanoparticle-protein interactions by fluorescamine labeling.

A rapid, high throughput fluorescence assay was designed to screen interactions between proteins and nanoparticles. The assay employs fluorescamine, a primary-amine specific fluorogenic dye, to label proteins. Because fluorescamine could specifically target the surface amines on proteins, a conformational change of the protein upon interaction with nanoparticles will result in a change in fluorescence. In the present study, the assay was applied to test the interactions between a selection of proteins and nanoparticles made of polystyrene, silica, or iron oxide. The particles were also different in their hydrodynamic diameter, synthesis procedure, or surface modification. Significant labeling differences were detected when the same protein incubated with different particles. Principal component analysis (PCA) on the collected fluorescence profiles revealed clear grouping effects of the particles based on their properties. The results prove that fluorescamine labeling is capable of detecting protein-nanoparticle interactions, and the resulting fluorescence profile is sensitive to differences in nanoparticle's physical properties. The assay can be carried out in a high-throughput manner, and is rapid with low operation cost. Thus, it is well suited for evaluating interactions between a larger number of proteins and nanoparticles. Such assessment can help to improve our understanding on the molecular basis that governs the biological behaviors of nanomaterials. It will also be useful for initial examination of the bioactivity and reproducibility of nanomaterials employed in biomedical fields.

[1]  Sidney Udenfriend,et al.  Fluorescamine: A Reagent for Assay of Amino Acids, Peptides, Proteins, and Primary Amines in the Picomole Range , 1972, Science.

[2]  V. Kolb-Bachofen,et al.  Coating particles with a block co-polymer (poloxamine-908) suppresses opsonization but permits the activity of dysopsonins in the serum. , 1993, Biochimica et biophysica acta.

[3]  Venyaminov SYu,et al.  Determination of protein tertiary structure class from circular dichroism spectra. , 1994, Analytical biochemistry.

[4]  M. Wahlgren,et al.  Structural Changes of T4 Lysozyme upon Adsorption to Silica Nanoparticles Measured by Circular Dichroism , 1995 .

[5]  John E. Coligan,et al.  Current Protocols in Protein Science , 1996 .

[6]  Werner Braun,et al.  Exact and efficient analytical calculation of the accessible surface areas and their gradients for macromolecules , 1998 .

[7]  R. Müller,et al.  'Stealth' corona-core nanoparticles surface modified by polyethylene glycol (PEG): influences of the corona (PEG chain length and surface density) and of the core composition on phagocytic uptake and plasma protein adsorption. , 2000, Colloids and surfaces. B, Biointerfaces.

[8]  I. Birlouez-Aragon,et al.  The FAST method, a rapid approach of the nutritional quality of heat-treated foods. , 2001, Die Nahrung.

[9]  E. Goormaghtigh,et al.  Protein concentration is not an absolute prerequisite for the determination of secondary structure from circular dichroism spectra: a new scaling method. , 2003, Analytical biochemistry.

[10]  T. Craig,et al.  Changes in structure and stability of calbindin-D(28K) upon calcium binding. , 2004, Analytical biochemistry.

[11]  Jonathan S Dordick,et al.  Silica nanoparticle size influences the structure and enzymatic activity of adsorbed lysozyme. , 2004, Langmuir : the ACS journal of surfaces and colloids.

[12]  V. Rotello,et al.  Monolayer-protected nanoparticle-protein interactions. , 2005, Current opinion in chemical biology.

[13]  N. C. Price,et al.  How to study proteins by circular dichroism. , 2005, Biochimica et biophysica acta.

[14]  Arezou A Ghazani,et al.  Determining the size and shape dependence of gold nanoparticle uptake into mammalian cells. , 2006, Nano letters.

[15]  A. Knight,et al.  A Comparison of Protein Quantitation Assays for Biopharmaceutical Applications , 2007, Molecular biotechnology.

[16]  Q. Xue,et al.  Preparation and self-assembly of carboxylic acid-functionalized silica. , 2007, Journal of colloid and interface science.

[17]  J. Schnekenburger,et al.  Not ready to use – overcoming pitfalls when dispersing nanoparticles in physiological media , 2008 .

[18]  J. Stockert,et al.  A mechanism for the fluorogenic reaction of amino groups with fluorescamine and MDPF. , 2008, Acta histochemica.

[19]  Kenneth A. Dawson,et al.  Nanoparticle size and surface properties determine the protein corona with possible implications for biological impacts , 2008, Proceedings of the National Academy of Sciences.

[20]  Lauren A Austin,et al.  Dynamic light scattering as a powerful tool for gold nanoparticle bioconjugation and biomolecular binding studies. , 2009, Analytical chemistry.

[21]  R. Albrecht,et al.  Toxicity assessments of multisized gold and silver nanoparticles in zebrafish embryos. , 2009, Small.

[22]  A. Perriman,et al.  Protein interfacial structure and nanotoxicology , 2009 .

[23]  M. Mann,et al.  Universal sample preparation method for proteome analysis , 2009, Nature Methods.

[24]  Bing Yan,et al.  Analytical strategies for detecting nanoparticle-protein interactions. , 2010, The Analyst.

[25]  Wenwan Zhong,et al.  Probing nanoparticle--protein interaction by capillary electrophoresis. , 2010, Analytical chemistry.

[26]  M. Mahmoudi,et al.  Protein-nanoparticle interactions: opportunities and challenges. , 2011, Chemical reviews.

[27]  Yang Chen,et al.  Fluorescent quantification of amino groups on silica nanoparticle surfaces , 2011, Analytical and bioanalytical chemistry.

[28]  Albert Duschl,et al.  Hardening of the nanoparticle-protein corona in metal (Au, Ag) and oxide (Fe3O4, CoO, and CeO2) nanoparticles. , 2011, Small.

[29]  Ronald J. Moore,et al.  Quantitative proteomics analysis of adsorbed plasma proteins classifies nanoparticles with different surface properties and size , 2011, Proteomics.

[30]  P. V. Asharani,et al.  Comparison of the toxicity of silver, gold and platinum nanoparticles in developing zebrafish embryos , 2011, Nanotoxicology.

[31]  Stefan Tenzer,et al.  Nanoparticle size is a critical physicochemical determinant of the human blood plasma corona: a comprehensive quantitative proteomic analysis. , 2011, ACS nano.

[32]  P. Guptasarma,et al.  N-Terminal sequencing by mass spectrometry through specific fluorescamine labeling of α-amino groups before tryptic digestion. , 2011, Analytical biochemistry.

[33]  Yi Cao,et al.  How do proteins unfold upon adsorption on nanoparticle surfaces? , 2012, Langmuir : the ACS journal of surfaces and colloids.

[34]  J. Dordick,et al.  Effect of gold nanoparticle structure on the conformation and function of adsorbed proteins. , 2012, Biomaterials.

[35]  Andrew Emili,et al.  Nanoparticle size and surface chemistry determine serum protein adsorption and macrophage uptake. , 2012, Journal of the American Chemical Society.

[36]  Wenwan Zhong,et al.  Impact of carrier fluid composition on recovery of nanoparticles and proteins in flow field flow fractionation. , 2012, Journal of chromatography. A.

[37]  Bengt Fadeel,et al.  Safety assessment of nanomaterials: implications for nanomedicine. , 2012, Journal of controlled release : official journal of the Controlled Release Society.

[38]  Albert Duschl,et al.  Interaction of nanoparticles with proteins: relation to bio-reactivity of the nanoparticle , 2013, Journal of Nanobiotechnology.

[39]  J. Driskell,et al.  Monitoring gold nanoparticle conjugation and analysis of biomolecular binding with nanoparticle tracking analysis (NTA) and dynamic light scattering (DLS). , 2013, The Analyst.

[40]  Wenwan Zhong,et al.  Dissociation-based screening of nanoparticle-protein interaction via flow field-flow fractionation. , 2013, Analytical chemistry.

[41]  R. von Klitzing,et al.  Impact of polymer shell on the formation and time evolution of nanoparticle-protein corona. , 2013, Colloids and surfaces. B, Biointerfaces.

[42]  Jingyuan Li,et al.  Revealing the binding structure of the protein corona on gold nanorods using synchrotron radiation-based techniques: understanding the reduced damage in cell membranes. , 2013, Journal of the American Chemical Society.

[43]  Teófilo Rojo,et al.  The challenge to relate the physicochemical properties of colloidal nanoparticles to their cytotoxicity. , 2013, Accounts of chemical research.

[44]  A. Ranz,et al.  Current Protocols in Protein Science , 2013 .

[45]  Huile Gao,et al.  The interaction of nanoparticles with plasma proteins and the consequent influence on nanoparticles behavior , 2014, Expert opinion on drug delivery.

[46]  Wenwan Zhong,et al.  Size and Surface Functionalization of Iron Oxide Nanoparticles Influence the Composition and Dynamic Nature of Their Protein Corona , 2014, ACS applied materials & interfaces.

[47]  Alexander Tropsha,et al.  Chemical basis of interactions between engineered nanoparticles and biological systems. , 2014, Chemical reviews.

[48]  Jacob Piehler,et al.  Spectroscopic techniques for monitoring protein interactions in living cells. , 2014, Current opinion in structural biology.