Isolation methods for particle protein corona complexes from protein-rich matrices

Böhmert, Linda; Voß, Linn; Stock, Valerie; Braeuning, Albert; Lampen, Alfonso; Sieg, Holger

doi:10.1039/c9na00537d

Cited by 64 publications

(75 citation statements)

References 220 publications

(307 reference statements)

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“…This interpretation of soft corona formation based on binding energy is in distinction from previous reports where the protein separable during centrifugation was associated with the presence of soft corona. 9 …”

Section: Results and Discussionmentioning

confidence: 99%

“… 7 Thus, any further interaction of NPs with biomolecules is mediated by the protein corona. 8 , 9 While most of the current studies focus on the biological activity of the protein within the corona, the mechanism of its formation and local orientation of proteins within the corona remain poorly understood. 10 , 11 This lack of understanding of protein corona characteristics is due to anisotropic shape and nonuniform distribution of chemical functional groups on the surface of protein molecules forming the corona.…”

Section: Introductionmentioning

confidence: 99%

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Adsorption of Myoglobin and Corona Formation on Silica Nanoparticles

et al. 2020

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The adsorption of proteins from aqueous medium leads to the formation of protein corona on nanoparticles. The formation of protein corona is governed by a complex interplay of protein−particle and protein−protein interactions, such as electrostatics, van der Waals, hydrophobic, hydrogen bonding, and solvation. The experimental parameters influencing these interactions, and thus governing the protein corona formation on nanoparticles, are currently poorly understood. This lack of understanding is due to the complexity in the surface charge distribution and anisotropic shape of the protein molecules. Here, we investigate the effect of pH and salinity on the characteristics of corona formed by myoglobin on silica nanoparticles. We experimentally measure and theoretically model the adsorption isotherms of myoglobin binding to silica nanoparticles. By combining adsorption studies with surface electrostatic mapping of myoglobin, we demonstrate that a monolayered hard corona is formed in low salinity dispersions, which transforms into a multilayered hard + soft corona upon the addition of salt. We attribute the observed changes in protein adsorption behavior with increasing pH and salinity to the change in electrostatic interactions and surface charge regulation effects. This study provides insights into the mechanism of protein adsorption and corona formation on nanoparticles, which would guide future studies on optimizing nanoparticle design for maximum functional benefits and minimum toxicity.

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Section: Results and Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Adsorption of Myoglobin and Corona Formation on Silica Nanoparticles

et al. 2020

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“…Deciphering intracellular trafficking pathways and mechanism (s) of endosomal escape will facilitate efforts to boost LNP potency [274][275][276][277]. Elucidating the nature and dynamic of the biomolecular corona formed on LNPs (following intravenous injection) and understanding its implications for biodistribution will be crucial to develop gene therapies beyond the liver [162][163][164]278,279]. Therefore, we expect that LNP-based gene therapies will be developed for indications beyond (ultra) rare diseases in the near future and increasingly become integrated in mainstream medicine.…”

Section: Future Perspectivesmentioning

confidence: 99%

Lipid nanoparticle technology for therapeutic gene regulation in the liver

Witzigmann

Kulkarni

Leung

et al. 2020

Advanced Drug Delivery Reviews

241

168

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Hereditary genetic disorders, cancer, and infectious diseases of the liver affect millions of people around the globe and are a major public health burden. Most contemporary treatments offer limited relief as they generally aim to alleviate disease symptoms. Targeting the root cause of diseases originating in the liver by regulating malfunctioning genes with nucleic acid-based drugs holds great promise as a therapeutic approach. However, employing nucleic acid therapeutics in vivo is challenging due to their unfavorable characteristics. Lipid nanoparticle (LNP) delivery technology is a revolutionary development that has enabled clinical translation of gene therapies. LNPs can deliver siRNA, mRNA, DNA, or gene-editing complexes, providing opportunities to treat hepatic diseases by silencing pathogenic genes, expressing therapeutic proteins, or correcting genetic defects. Here we discuss the state-of-the-art LNP technology for hepatic gene therapy including formulation design parameters, production methods, preclinical development and clinical translation.

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“…Many factors affect the PC composition; potential factors influencing the PC include characteristics of the NPs (i.e., size, charge, and surface engineering), characteristics of the protein (i.e., molecular weight, isoelectric points, structure and folding), characteristics of the interaction (i.e., time, temperature, concentration), and the type of biological fluid (i.e., plasma, urine, tissue lysate, etc.) (Bohmert et al 2020;Tenzer et al 2013;Deng et al 2009;Vogt et al 2015;Hadjidemetriou et al 2015Hadjidemetriou et al , 2016Aggarwal et al 2009). The PC composition is also modulated by the separation technique used to recover NPs from the NP-biological fluid mixture, i.e., centrifugation, ultracentrifugation, magnetism, or chromatography (Bohmert et al 2020).…”

Section: Introductionmentioning

confidence: 99%

“…(Bohmert et al 2020;Tenzer et al 2013;Deng et al 2009;Vogt et al 2015;Hadjidemetriou et al 2015Hadjidemetriou et al , 2016Aggarwal et al 2009). The PC composition is also modulated by the separation technique used to recover NPs from the NP-biological fluid mixture, i.e., centrifugation, ultracentrifugation, magnetism, or chromatography (Bohmert et al 2020). Similarly, pre-processing conditions may impact the PC composition.…”

Section: Introductionmentioning

confidence: 99%

Experimental conditions influence the formation and composition of the corona around gold nanoparticles

et al. 2021

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Background Ovarian cancer is one of the deadliest gynecological malignancies. While the overall survival of ovarian cancer patients has slightly improved in recent years in the developed world, it remains clinically challenging due to its frequent late diagnosis and the lack of reliable diagnostic and/or prognostic markers. The aim of this study was to identify potential new molecular target proteins (NMTPs) responsible for the poor outcomes. When nanoparticles (NP) are exposed to biological fluids, a protein coat, termed the protein corona (PC), forms around the NP, and the PC represents a tool to identify NMTPs. This study investigates the influence of pre-processing conditions, such as lysis conditions and serum/plasma treatment, on the PC composition and the resulting identification of NMTPs. Results Using gel electrophoresis, pre-processing conditions, including cell-lysis techniques and enrichment of low-abundance proteins (LAPs) by immunocentrifugation of serum/plasma, were shown to alter the relative amounts and compositions of proteins. PCs formed when 20 nm gold-NPs (GNPs) were incubated with lysate proteins from either RIPA- or urea lysis. Proteomic analysis of these PCs showed 2–22-fold enrichment of NMTPs in PCs from urea lysates as compared to RIPA lysates. Enriched NMTPs were then classified as cellular components, biological and molecular functions-associated proteins. The impact of enriched LAPs (eLAPs) on both PC composition and NMTP identification was shown by comparative proteomic analysis of original plasma, eLAPs, and PCs derived from eLAPs; eLAPs-PCs enhanced the abundance of NMTPs approximately 13%. Several NMTPs, including gasdermin-B, dermcidin, and kallistatin, were identified by this method demonstrating the potential use of this PC approach for molecular target discovery. Conclusion The current study showed that the pre-processing conditions modulate PC composition and can be used to enhance identification of NMTPs.

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Isolation methods for particle protein corona complexes from protein-rich matrices

Abstract: This review compares and critically discusses the most widely used analysis techniques for protein corona investigations: centrifugation, magnetism and chromatography.

Cited by 64 publications

References 220 publications

Adsorption of Myoglobin and Corona Formation on Silica Nanoparticles

Adsorption of Myoglobin and Corona Formation on Silica Nanoparticles

Lipid nanoparticle technology for therapeutic gene regulation in the liver

Experimental conditions influence the formation and composition of the corona around gold nanoparticles

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