scholarly journals Protein corona formation around biocatalytic nanomotors unveiled by STORM

2021 ◽  
Author(s):  
Tania Patino ◽  
Joaquin Llacer-Wintle ◽  
Silvia Pujals ◽  
Lorenzo Albertazzi ◽  
Samuel Sánchez
Keyword(s):  
Serum Proteins ◽  
Biological Fluids ◽  
Protein Corona ◽  
General Interest ◽  
Biological Media ◽  
Enhanced Diffusion ◽  
Stochastic Optical Reconstruction Microscopy ◽  
Corona Formation ◽  
Active Nanoparticles ◽  
Optical Reconstruction

The interaction of nanoparticles with biological media is a topic of general interest for drug delivery systems and among those for active nanoparticles, also called nanomotors. Herein, we report the use of super resolu-tion microscopy, in particular stochastic optical reconstruction microscopy (STORM), to characterize the formation of protein corona around active enzyme-powered nanomotors. First, we characterize the distribu-tion and number of enzymes on nano-sized particles and characterized their motion capabilities. Then, we incubated the nanomotors with fluorescently labelled serum proteins. Interestingly, we observed a signifi-cant decrease of protein corona formation (20 %) and different composition, which was studied by a proteo-mic analysis. Moreover, motion was not hindered, as nanomotors displayed an enhanced diffusion regardless of protein corona. Elucidating how active particles interact with biological media and maintain their self-propulsion after protein corona formation will pave the way of the use these systems in complex biological fluids in biomedicine.

Silver nanoparticles in complex biological media: assessment of colloidal stability and protein corona formation

2016 ◽  
Vol 18 (8) ◽  
Author(s):  
Simona Argentiere ◽  
Claudia Cella ◽  
Maura Cesaria ◽  
Paolo Milani ◽  
Cristina Lenardi
Keyword(s):  
Silver Nanoparticles ◽  
Colloidal Stability ◽  
Protein Corona ◽  
Biological Media ◽  
Corona Formation

scholarly journals Gold Nanostars with Reduced Fouling Facilitate Small Molecule Detection in the Presence of Protein

Nanomaterials ◽  
2021 ◽  
Vol 11 (10) ◽  
pp. 2565
Author(s):  
Anastasiia Tukova ◽  
Inga Christine Kuschnerus ◽  
Alfonso Garcia-Bennett ◽  
Yuling Wang ◽  
Alison Rodger
Keyword(s):  
Gold Nanoparticles ◽  
Small Molecule ◽  
Protein Corona ◽  
Surface Enhanced Raman Spectroscopy ◽  
Biological Media ◽  
Gold Nanostars ◽  
Cellular Environment ◽  
Vis Spectroscopy ◽  
Corona Formation

Gold nanoparticles have the potential to be used in biomedical applications from diagnostics to drug delivery. However, interactions of gold nanoparticles with different biomolecules in the cellular environment result in the formation of a “protein corona”—a layer of protein formed around a nanoparticle, which induces changes in the properties of nanoparticles. In this work we developed methods to reproducibly synthesize spheroidal and star-shaped gold nanoparticles, and carried out a physico-chemical characterization of synthesized anionic gold nanospheroids and gold nanostars through transmission electron microscopy (TEM), dynamic light scattering (DLS), zeta potential (ZP), nanoparticles tracking analysis (NTA), ultraviolet-visible (UV–Vis) spectroscopy and estimates of surface-enhanced Raman spectroscopy (SERS) signal enhancement ability. We analyzed how they interact with proteins after pre-incubation with bovine serum albumin (BSA) via UV–Vis, DLS, ZP, NTA, SERS, cryogenic TEM (cryo-TEM) and circular dichroism (CD) spectroscopy. The tests demonstrated that the protein adsorption on the particles’ surfaces was different for spheroidal and star shaped particles. In our experiments, star shaped particles limited the protein corona formation at SERS “hot spots”. This benefits the small-molecule sensing of nanostars in biological media. This work adds more understanding about protein corona formation on gold nanoparticles of different shapes in biological media, and therefore guides design of particles for studies in vitro and in vivo.

scholarly journals Time evolution of PEG-shedding and serum protein coronation determines the cell uptake kinetics and delivery of lipid nanoparticle formulated mRNA

2021 ◽  
Author(s):  
Audrey Gallud ◽  
Michael J Munson ◽  
Kai Liu ◽  
Alexander Idstrom ◽  
Hanna MG Barriga ◽  
...  
Keyword(s):  
Serum Proteins ◽  
Short Pulse ◽  
High Surface ◽  
Protein Corona ◽  
Cell Uptake ◽  
Lipid Nanoparticle ◽  
Nanoparticle Imaging ◽  
Early Endosomes ◽  
Corona Formation ◽  
Model Protein

Development of efficient lipid nanoparticle (LNP) vectors remains a major challenge towards broad clinical translation of RNA therapeutics. New lipids will be required, but also better understanding LNP interactions with the biological environment. Herein, we model protein corona formation on PEG-ylated DLin-MC3-DMA LNPs and identify time-dependent maturation steps that critically unlock their cellular uptake and mRNA delivery. Uptake requires active serum proteins and precedes after a significant (~2 hours) lag-time, which we show can be eliminated by pre-incubating LNPs for 3-4 hours in serum-containing media. This indicates an important role of protein corona maturation for the pharmacokinetic effects of these LNPs. We show, using single-nanoparticle imaging, NMR diffusometry, SANS, and proteomics, that the LNPs, upon serum exposure, undergo rapid PEG-shedding (~30 minutes), followed by a slower rearrangement of the adsorbed protein layer. The PEG-shedding coincides in time with high surface abundance of Apolipoprotein A-II, whereas the LNPs preferentially bind Apolipoprotein E when their maximum uptake-competent state is reached. Finally, we show that pre-incubation of the LNPs enables rapid uptake and allows pulse-chase video-microscopy colocalization experiments with sufficiently short pulse durations to gain improved mechanistic understanding of how intracellular trafficking events determine delivery efficacy, emphasizing early endosomes as important delivery-mediating compartments.

scholarly journals Understanding the Lipid and Protein Corona Formation on Different Sized Polymeric Nanoparticles

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Tânia Lima ◽  
Katja Bernfur ◽  
Manuel Vilanova ◽  
Tommy Cedervall
Keyword(s):  
Serum Proteins ◽  
Biological Fluids ◽  
Complex Structure ◽  
Biological Properties ◽  
Host Cells ◽  
Mouse Serum ◽  
Corona Formation ◽  
The Common ◽  
The Impact

AbstractWhen in contact with biological fluids, nanoparticles dynamically absorb biomolecules like proteins and lipids onto their surface, forming a “corona”. This biocorona is a dynamic and complex structure that determines how host cells respond to nanoparticles. Despite the common use of mouse models in pre-clinical and toxicological experiments, the impact of corona formed in mouse serum on the biophysical and biological properties of different size NP has not been thoroughly explored. Furthering the knowledge on the corona formed on NP exposed to mouse serum proteins can help in understanding what role it might have in in vivo studies at systemic, tissue, and cellular levels. To investigate biocorona formation, different sized polystyrene NP were exposed to mouse serum. Our data show a size- and time-dependent protein and lipid corona formation. Several proteins were identified and apolipoproteins were by far the most common group on the NPs surfaces. Moreover, we observed that cholesterol and triglycerides effectively bind to NP emphasizing that proteins are not the only biomolecules with high-affinity binding to nanomaterial surfaces. These results highlight that further knowledge on NP interactions with mouse serum is necessary regarding the common use of this model to predict the in vivo efficiency of NP.

scholarly journals Blinking statistics and molecular counting in direct stochastic reconstruction microscopy (dSTORM)

2021 ◽  
Author(s):  
Lekha Patel ◽  
David Williamson ◽  
Dylan M Owen ◽  
Edward A K Cohen
Keyword(s):  
Probability Distribution ◽  
Single Molecule ◽  
Immunological Synapse ◽  
Training Data ◽  
Supplementary Information ◽  
Cellular Structures ◽  
Exact Probability ◽  
Stochastic Optical Reconstruction Microscopy ◽  
Exact Probability Distribution ◽  
Optical Reconstruction

Abstract Motivation Many recent advancements in single-molecule localization microscopy exploit the stochastic photoswitching of fluorophores to reveal complex cellular structures beyond the classical diffraction limit. However, this same stochasticity makes counting the number of molecules to high precision extremely challenging, preventing key insight into the cellular structures and processes under observation. Results Modelling the photoswitching behaviour of a fluorophore as an unobserved continuous time Markov process transitioning between a single fluorescent and multiple dark states, and fully mitigating for missed blinks and false positives, we present a method for computing the exact probability distribution for the number of observed localizations from a single photoswitching fluorophore. This is then extended to provide the probability distribution for the number of localizations in a direct stochastic optical reconstruction microscopy experiment involving an arbitrary number of molecules. We demonstrate that when training data are available to estimate photoswitching rates, the unknown number of molecules can be accurately recovered from the posterior mode of the number of molecules given the number of localizations. Finally, we demonstrate the method on experimental data by quantifying the number of adapter protein linker for activation of T cells on the cell surface of the T-cell immunological synapse. Availability and implementation Software and data available at https://github.com/lp1611/mol_count_dstorm. Supplementary information Supplementary data are available at Bioinformatics online.

scholarly journals Molecular organization of mammalian meiotic chromosome axis revealed by expansion STORM microscopy

2019 ◽  
Vol 116 (37) ◽  
pp. 18423-18428 ◽  
Author(s):  
Huizhong Xu ◽  
Zhisong Tong ◽  
Qing Ye ◽  
Tengqian Sun ◽  
Zhenmin Hong ◽  
...  
Keyword(s):  
Meiotic Chromosome ◽  
Molecular Organization ◽  
Homologous Chromosomes ◽  
Meiotic Chromosomes ◽  
C Terminus ◽  
Stochastic Optical Reconstruction Microscopy ◽  
Optical Reconstruction ◽  
20 Nm ◽  
Chromosome Axis

During prophase I of meiosis, chromosomes become organized as loop arrays around the proteinaceous chromosome axis. As homologous chromosomes physically pair and recombine, the chromosome axis is integrated into the tripartite synaptonemal complex (SC) as this structure’s lateral elements (LEs). While the components of the mammalian chromosome axis/LE—including meiosis-specific cohesin complexes, the axial element proteins SYCP3 and SYCP2, and the HORMA domain proteins HORMAD1 and HORMAD2—are known, the molecular organization of these components within the axis is poorly understood. Here, using expansion microscopy coupled with 2-color stochastic optical reconstruction microscopy (STORM) imaging (ExSTORM), we address these issues in mouse spermatocytes at a resolution of 10 to 20 nm. Our data show that SYCP3 and the SYCP2 C terminus, which are known to form filaments in vitro, form a compact core around which cohesin complexes, HORMADs, and the N terminus of SYCP2 are arrayed. Overall, our study provides a detailed structural view of the meiotic chromosome axis, a key organizational and regulatory component of meiotic chromosomes.

Sign in / Sign up

Export Citation Format

Share Document