In situ characterization of nanoparticle biomolecular interactions in complex biological media by flow cytometry

Giudice, Cristina Lo; Herda, Luciana M.; Polo, Ester; Dawson, Kenneth A.

doi:10.1038/ncomms13475

Cited by 146 publications

(118 citation statements)

References 52 publications

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“…While there is extensive literature dealing with the subject of hard protein corona, only limited analytic methods used to study the soft protein corona are available. 21 Consequently, determining the biological relevance of the soft corona has been slowed down. 22 Hence, for in vivo application of nanocarriers it is essential to concentrate on studying the interactions of nanoparticles within a given protein source (serum, plasma) and to analyze their aggregation behavior in such complex surroundings as this can highly affect the biodistribution.…”

Section: Resultsmentioning

confidence: 99%

Visualization of the protein corona: towards a biomolecular understanding of nanoparticle-cell-interactions

et al. 2017

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The use of nanocarriers in biology and medicine is complicated by the current need to understand how nanoparticles interact in complex biological surroundings. When nanocarriers come into contact with serum, proteins immediately adsorb onto their surface, forming a protein corona which defines their biological identity. Although the composition of the protein corona has been widely determined by proteomics, its morphology still remains unclear. In this study we show for the first time the morphology of the protein corona using transmission electron microscopy. We are able to demonstrate that the protein corona is not, as commonly supposed, a dense, layered shell coating the nanoparticle, but an undefined, loose network of proteins. Additionally, we are now able to visualize and discriminate between the soft and hard corona using centrifugation-based separation techniques together with proteomic characterization. The protein composition of the ∼15 nm hard corona strongly depends on the surface chemistry of the respective nanomaterial, thus further affecting cellular uptake and intracellular trafficking. Large diameter protein corona resulting from pre-incubation with soft corona or Apo-A1 inhibits cellular uptake, confirming the stealth-effect mechanism. In summary, the knowledge on protein corona formation, composition and morphology is essential to design therapeutic effective nanoparticle systems.

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Section: Resultsmentioning

confidence: 99%

Visualization of the protein corona: towards a biomolecular understanding of nanoparticle-cell-interactions

et al. 2017

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“…Our suggestion is supported by recent literature showing that LDL-enriched NPs present functional motifs that allow recognition by LDL receptor 38 . Our future biological understanding of how NPs interact with cells will confirm these conclusions more in depth by benefiting from a detailed knowledge of the arrangements of functional motifs of BC components at the nanoscale 39–41 .…”

Section: Resultsmentioning

confidence: 81%

An apolipoprotein-enriched biomolecular corona switches the cellular uptake mechanism and trafficking pathway of lipid nanoparticles

et al. 2017

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Following exposure to biological milieus (e.g. after systemic administration), nanoparticles (NPs) get covered by an outer biomolecular corona (BC) that defines many of their biological outcomes, such as the elicited immune response, biodistribution, and targeting capabilities. In spite of this, the BC role in regulating the cellular uptake and the subcellular trafficking properties of NPs has remained elusive. Here, we tackle this issue by employing multicomponent (MC) lipid NPs, human plasma (HP) and HeLa cells as models for nanoformulation, biological fluid, and target cell (respectively). By confocal fluorescence microscopy experiments and image correlation analyses, we quantitatively demonstrate that the BC promotes a neat switch of cell entry mechanism and subsequent intracellular trafficking, from macropinocytosis to clathrin-dependent endocytosis. Nano liquid chromatography tandem mass spectrometry identifies Apolipoproteins as the most abundant components of the BC tested here. Interestingly, this class of proteins target the LDL receptors, which are overexpressed in clathrin-enriched membrane domains. Our results highlight the crucial role of BC as an intrinsic trigger of specific NP-cell interactions and biological responses and set the basis for a rational exploitation of the BC for targeted delivery.

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“…Flow cytometry (FC) was used to evaluate different parameters among the different nMB preparations (Figure 1d; Figure S8, Supporting Information);29 that is, the nMB's size distribution (Figure 1d, panel i, Side Scattering), cargo loading (Figure 1d, panel ii, Cargo), the presence of lipids (Figure 1d, panel iii, Cell Membrane), and the presence and correct orientation of some cell adhesion proteins (Figure 1d, panel iv, Transmembrane Protein), as a reporter for the cadherin‐family proteins by using a fluorescent‐labeled Ab. The immunolabeling detection of the extracellular domain of the cadherins indicated a right‐side‐out membrane orientation on the synthesized nMBs.…”

Section: Resultsmentioning

confidence: 99%

Plasmonic Cell‐Derived Nanocomposites for Light‐Controlled Cargo Release inside Living Cells

et al. 2020

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Translating the potential of thermoplasmonics to cell‐derived nanomaterials offers exciting opportunities to fabricate beyond state‐of‐art artificial biomimetic nanocomposites that upon illumination perform active tasks such as delivery of cargo in complex, dynamic media such as the cytosol of cells. Cell‐derived nanoparticles have shown stunning potential to implement cell‐specific functions, such as long blood circulation or targeting capabilities, into advanced drug delivery nanosystems. The biomimicry nanotechnology has now advanced to offer new and exciting opportunities to improve the commonly poor in vivo performance of most current nanomedicines, including evading the immune system and targeting specific tissues such as tumors, the latest remaining among the most wanted breakthroughs in nanomedicine. However, the use of cell‐derived nanocomposites as stimulus‐controlled drug delivery agents remains virtually unexplored. This study reports the fabrication of a plasmonic cell‐derived nanocomposite by integrating near‐infrared active gold nanorods in its structure. As a proof of concept, the plasmonic nanomembranes are loaded with cell non‐permeant antibodies, which upon near‐infrared stimulation can be released from the plasmonic nanomembranes into the cytosol of living cells, without impairing cell viability or the antibodies' function. These results set the stage for the development of photoactive cell‐derived nanocarriers, which in addition to cell‐specific functions promise straightforward access to spatiotemporal‐controlled intracellular delivery of antibodies.

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In situ characterization of nanoparticle biomolecular interactions in complex biological media by flow cytometry

Cited by 146 publications

References 52 publications

Visualization of the protein corona: towards a biomolecular understanding of nanoparticle-cell-interactions

Visualization of the protein corona: towards a biomolecular understanding of nanoparticle-cell-interactions

An apolipoprotein-enriched biomolecular corona switches the cellular uptake mechanism and trafficking pathway of lipid nanoparticles

Plasmonic Cell‐Derived Nanocomposites for Light‐Controlled Cargo Release inside Living Cells

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