Quantitative analysis of the protein corona on FePt nanoparticles formed by transferrin binding

Nanoparticles are finding a rapidly expanding range of applications in research and technology, finally entering our daily life in medical, cosmetic or food products. Their ability to invade all regions of an organism including cells and cellular organelles offers new strategies for medical diagnosis and therapy (nanomedicine), but their safe use requires a deep knowledge about their interactions with biological systems at the molecular level. Upon incorporation, nanoparticles are exposed to biological fluids from which they adsorb proteins and other biomolecules to form a ‘protein corona’. These nanoparticle–protein interactions are still poorly understood and quantitative studies to characterize them remain scarce. Here we have quantitatively analysed the adsorption of human transferrin onto small (radius approx. 5 nm) polymer-coated FePt nanoparticles by using fluorescence correlation spectroscopy. Transferrin binds to the negatively charged nanoparticles with an affinity of approximately 26 µM in a cooperative fashion and forms a monolayer with a thickness of 7 nm. By using confocal fluorescence microscopy, we have observed that the uptake of FePt nanoparticles by HeLa cells is suppressed by the protein corona compared with the bare nanoparticles.

[1]  L. Manna,et al.  Synthesis and perspectives of complex crystalline nano‐structures , 2006 .

[2]  D. C. Carter,et al.  Atomic structure and chemistry of human serum albumin , 1993, Nature.

[3]  T C Fisher,et al.  The hydrodynamic radii of macromolecules and their effect on red blood cell aggregation. , 2004, Biophysical journal.

[4]  Wolfgang J Parak,et al.  Combined atomic force microscopy and optical microscopy measurements as a method to investigate particle uptake by cells. , 2006, Small.

[5]  K. Landfester,et al.  Uptake of functionalized, fluorescent-labeled polymeric particles in different cell lines and stem cells. , 2006, Biomaterials.

[6]  R. Rigler,et al.  Fluorescence correlation spectroscopy , 2001 .

[7]  V. Colvin The potential environmental impact of engineered nanomaterials , 2003, Nature Biotechnology.

[8]  R. Levy,et al.  Arterial uptake of biodegradable nanoparticles: effect of surface modifications. , 1998, Journal of pharmaceutical sciences.

[9]  Junaed Sattar Snakes , Shapes and Gradient Vector Flow , 2022 .

[10]  Russell J Mumper,et al.  Comparison of cell uptake, biodistribution and tumor retention of folate-coated and PEG-coated gadolinium nanoparticles in tumor-bearing mice. , 2004, Journal of controlled release : official journal of the Controlled Release Society.

[11]  J. Hamers,et al.  [Methods and techniques]. , 1997, Verpleegkunde.

[12]  M. Bock,et al.  Physical and Biological Characterization of Superparamagnetic Iron Oxide- and Ultrasmall Superparamagnetic Iron Oxide-Labeled Cells: A Comparison , 2005, Investigative radiology.

[13]  W. Shen,et al.  Mechanisms of TfR-mediated transcytosis and sorting in epithelial cells and applications toward drug delivery. , 2003, Advanced drug delivery reviews.

[14]  J. Lakowicz Topics in fluorescence spectroscopy , 2002 .

[15]  Sara Linse,et al.  Detailed identification of plasma proteins adsorbed on copolymer nanoparticles. , 2007, Angewandte Chemie.

[16]  M. Walker,et al.  Mammalian class I myosin, Myo1b, is monomeric and cross-links actin filaments as determined by hydrodynamic studies and electron microscopy. , 2005, Biophysical journal.

[17]  Hongzhe Sun,et al.  Targeted Drug Delivery via the Transferrin Receptor-Mediated Endocytosis Pathway , 2002, Pharmacological Reviews.

[18]  C. Röcker,et al.  Fluctuation correlation spectroscopy for the advanced physics laboratory , 2005 .

[19]  Wolfgang J Parak,et al.  A quantitative fluorescence study of protein monolayer formation on colloidal nanoparticles. , 2009, Nature nanotechnology.

[20]  A Ciechanover,et al.  Kinetics of internalization and recycling of transferrin and the transferrin receptor in a human hepatoma cell line. Effect of lysosomotropic agents. , 1983, The Journal of biological chemistry.

[21]  Sara Linse,et al.  Understanding the nanoparticle–protein corona using methods to quantify exchange rates and affinities of proteins for nanoparticles , 2007, Proceedings of the National Academy of Sciences.

[22]  M. Ferrer,et al.  The conformation of serum albumin in solution: a combined phosphorescence depolarization-hydrodynamic modeling study. , 2001, Biophysical journal.

[23]  A. Barendregt,et al.  Synthesis and perspectives for the future , 2009 .

[24]  Mathias Brust,et al.  Uptake and intracellular fate of surface-modified gold nanoparticles. , 2008, ACS nano.

[25]  S. Schürch,et al.  Interaction of fine particles and nanoparticles with red blood cells visualized with advanced microscopic techniques. , 2006, Environmental science & technology.

[26]  Pablo D. Jadzinsky,et al.  Structure of a Thiol Monolayer-Protected Gold Nanoparticle at 1.1 Å Resolution , 2007, Science.

[27]  Elliot L. Elson,et al.  Fluorescence correlation spectroscopy : theory and applications , 2001 .

[28]  Sandra L. Schmid,et al.  Regulated portals of entry into the cell , 2003, Nature.

[29]  Walter H. Chang,et al.  Design of an amphiphilic polymer for nanoparticle coating and functionalization. , 2008, Small.

[30]  G Ulrich Nienhaus,et al.  Confocal optics microscopy for biochemical and cellular high-throughput screening. , 2003, Drug discovery today.

[31]  N. Thompson,et al.  Fluorescence Correlation Spectroscopy , 2002 .

[32]  Thomas Dertinger,et al.  Two-focus fluorescence correlation spectroscopy: a new tool for accurate and absolute diffusion measurements. , 2007, Chemphyschem : a European journal of chemical physics and physical chemistry.

[33]  H. Jhoti,et al.  Molecular structure of serum transferrin at 3. 3-A resolution , 1988 .

[34]  I. Zuhorn,et al.  Size-dependent internalization of particles via the pathways of clathrin- and caveolae-mediated endocytosis. , 2004, The Biochemical journal.

[35]  A. P. Alivisatos,et al.  Shape control and applications of nanocrystals , 2003, Philosophical Transactions of the Royal Society of London. Series A: Mathematical, Physical and Engineering Sciences.

[36]  M. Gunzburg,et al.  Experimental determination of quantum dot size distributions, ligand packing densities, and bioconjugation using analytical ultracentrifugation. , 2008, Nano letters.

[37]  G Ulrich Nienhaus,et al.  A far-red fluorescent protein with fast maturation and reduced oligomerization tendency from Entacmaea quadricolor (Anthozoa, Actinaria) , 2002, Proceedings of the National Academy of Sciences of the United States of America.

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

[39]  Katharina Landfester,et al.  Synthesis and biomedical applications of functionalized fluorescent and magnetic dual reporter nanoparticles as obtained in the miniemulsion process , 2006 .

[40]  Hongzhe Sun,et al.  Transferrin-mediated gold nanoparticle cellular uptake. , 2005, Bioconjugate chemistry.

[41]  D. Lamb,et al.  Sensitivity enhancement in fluorescence correlation spectroscopy of multiple species using time-gated detection. , 2000, Biophysical journal.

[42]  G Ulrich Nienhaus,et al.  Photodynamics of red fluorescent proteins studied by fluorescence correlation spectroscopy. , 2004, Biophysical journal.

[43]  Jeff W M Bulte,et al.  Intracytoplasmic tagging of cells with ferumoxides and transfection agent for cellular magnetic resonance imaging after cell transplantation: methods and techniques , 2003, Transplantation.

[44]  Richard O. Duda,et al.  Pattern classification and scene analysis , 1974, A Wiley-Interscience publication.

[45]  Peter Burgherr,et al.  Synthesis and Perspectives , 2003 .

[46]  W. Webb,et al.  Fluorescence correlation spectroscopy: diagnostics for sparse molecules. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[47]  Thomas Walz,et al.  Structure of the Human Transferrin Receptor-Transferrin Complex , 2004, Cell.