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Concentration and composition of the protein corona as a function of incubation time and serum concentration

ABSTRACT: We describe the concentration and composition of the protein corona as a function of incubation time (30 min and 1 day) and the concentration of fetal bovine serum (FBS) used to form the corona (5%, 10%, and 100%) on 200 nm carboxylate-modified magnetic NPs, using semi-automated processes.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Bos Taurus (bovine)

TISSUE(S): Blood Serum

SUBMITTER: Karsten Poulsen

LAB HEAD: Christine Payne

PROVIDER: PXD035811 | Pride | 2023-03-21

REPOSITORIES: pride

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			Action	DRS
	ID65137_01_FSN20357_5611_061421.index	Other
	ID65137_01_FSN20357_5611_061421.raw	Raw
	ID66204_01_OTIT_FSN20357_5611_072721.index	Other
	ID66204_01_OTIT_FSN20357_5611_072721.raw	Raw
	ID67501_01_FSN20560_5611_102621.index	Other

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Publications

Concentration and composition of the protein corona as a function of incubation time and serum concentration: an automated approach to the protein corona.

Poulsen Karsten M KM Payne Christine K CK

Analytical and bioanalytical chemistry 20220826 24

Nanoparticles in contact with proteins form a "corona" of proteins adsorbed on the nanoparticle surface. Subsequent biological responses are then mediated by the adsorbed proteins rather than the bare nanoparticles. The use of nanoparticles as nanomedicines and biosensors would be greatly improved if researchers were able to predict which specific proteins will adsorb on a nanoparticle surface. We use a recently developed automated workflow with a liquid handling robot and low-cost proteomics to ...[more]

PMID: 36018335

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Interaction of TiO2 nanoparticles with lung fluid proteins and the resulting macrophage inflammatory response

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Mass spectrometry-based top-down proteomics in nanomedicine: proteoform-specific measurement of protein corona

Project description:Conventional mass spectrometry (MS)-based bottom-up proteomics (BUP) analysis of protein corona [i.e., an evolving layer of biomolecules, mostly proteins, formed on the surface of nanoparticles (NPs) during their interactions with biomolecular fluids] enabled nanomedicine community to partly identify the biological identity of NPs. Such an approach, however, fails pinpoint the specific proteoforms—distinct molecular variants of proteins, which is essential for prediction of the biological fate and pharmacokinetics of nanomedicines. Recognizing this limitation, this study pioneers a robust and reproducible MS-based top-down proteomics (TDP) technique for precisely characterizing proteoforms in the protein corona. Our TDP approach has successfully identified hundreds of proteoforms in the protein corona of polystyrene NPs, ranging from 3-70 kDa, revealing over 20 protein biomarkers with combinations of post-translational modifications, signal peptide cleavages, and/or truncations—details that BUP could not fully discern. This advancement in MS-based TDP offers a more comprehensive and exact characterization of NP protein coronas, deepening our understanding of NPs' biological identities and potentially revolutionizing the field of nanomedicine.

2024-10-15 | PXD050779 | Pride

Deep Plasma Proteome Profiling by Modulating Single Nanoparticle Protein Corona with Small Molecules

Project description:The protein corona, a dynamic biomolecular layer that forms on nanoparticle (NP) surfaces upon exposure to biological fluids is emerging as a valuable diagnostic tool for improving plasma proteome coverage analyzed by liquid chromatography-mass spectrometry (LC-MS/MS). Here, we show that spiking phosphatidylcholine into plasma can induce diverse protein corona patterns on otherwise identical NPs, significantly enhancing the depth of plasma proteome profiling. This significant achievement is made utilizing only a single NP type and one small molecule to analyze a single plasma sample, setting a new standard in proteomic depth of the plasma sample. Given the critical role of plasma proteomics in biomarker discovery and disease monitoring, we anticipate widespread adoption of this methodology for identification and clinical translation of proteomic biomarkers into FDA approved diagnostics.

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Mechanism of protein corona and surface modification of MSNs affecting the tissue distribution

Project description:Once NPs were administered into the physiological condition, several serum proteins could adsorb rapidly onto the NPs to form a protein layer (also called protein corona). The effect of protein corona surrounding the NPs depends on several parameters such as size, charge and other surface properties of NPs, which could contribute a great deal of influence in determining the NP's fate.

2022-08-12 | PXD033783 | Pride

STRT RNAseq of THP-1 monocytic cells exposed to surface modified-gold nanoparticles

Project description:Transcriptome comparison to assess the responses of human monocytic cells (THP-1) to gold-nanoparticles (Au-NPs) of two different diameter sizes (5 or 20 nm) with different surface functional groups, i.e., alkylammonium bromide, alkyl sodium carboxylate, or poly(ethylene glycol) (PEG)-terminated thiolate Au-NPs.

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2022-08-12 | PXD026615 | Pride

Influence of Albumin Concentration on Surface Characteristics and Cellular Responses in the Pre-Incubation of Multi-Walled Carbon Nanotubes

Project description:Due to the characteristics of multi-walled carbon nanotubes (MWCNTs) to aggregate and bind non-specifically, attempts have been made to inhibit their aggregation and increase their dispersion stability for biomedical applications by coating with bovine serum albumin (BSA). However, BSA concentration has not been studied in detail in terms of cellular response. To investigate the surface proteins formed in 10% fetal bovine serum cell culture media depending on the BSA concentration used to stabilize MWCNTs and to correlate the cellular responses with BSA concentration, we performed nanoflow liquid chromatography–electrospray ionization–tandem mass spectrometry analysis to quantify proteins of the protein corona (PC) and of A549 cells. Results showed that increasing the concentration of pre-incubated BSA changed the composition ratio of the proteins of the PCs adsorbed on the MWCNTs and also decreased the cellular uptake of A549 cells treated with BSA-coated MWCNTs. In the proteomic analysis, proteins related to the ribosome, ribosome biogenesis in eukaryotes, and oxidative phosphorylation pathways were downregulated in the low concentration BSA-treated cell groups, while proteins related to the mRNA surveillance pathway were downregulated in the high-BSA-concentration cell groups. Moreover, circular dichroism and Fourier-transform infrared spectroscopy results showed that the α-helix structure, a secondary structure of the adsorbed proteins, increased as a function of BSA concentration. Taken together, BSA concentration affects the adsorption and secondary structure of PCs on MWCNTs, which in turns affect their intracellular behavior. Our results may provide insight for the design of modification methods to increase the biocompatibility of MWCNTs.

2024-10-21 | PXD052226 | Pride

Human granulosa

Project description:We are human embryologists from center for reproductive medicinel of Anhui Provincial Hospital Affiliated to Anhui Medical University, and we have the expertise to do all that properly in humans. By deep sequencing, the current experiment determined the miRNA profile of two intrafollicular somatic cell types: CRCs and COCs, isolated from women undergoing controlled ovarian stimulation and in vitro fertilization treatment. Ovarian follicles, which are a densely-packed shell of granulosa cells that contains an immature or mature oocyte, are above all responsible for the development, maturation, and release of mature egg for fertilization. They are also responsible for synthesizing and secreting hormones that are essential for follicular development, menstrual and estrous cycle, maintenance of the reproductive tracts and their functions, development of female secondary sex characteristics, and metabolism. During folliculogenesis, ovarian granulosa cells surrounding the oocyte differentiate into mural granulosa cells, involved in gonadal steroidogenesis, and into cumulus cells, which are ovulated with the oocyte at ovulation. These cumulus cells derive from the same population of early follicles, but differentiate into two distinct groups of cells: 1) Those directly lie on the zona pellucida are composed of the so called corona radiata cells.(CRCs) 2) The other group surrounds the CRCs and consists of more numerous cells, forming the so called cumulus oophorus cells (COCs). In the present study, we described the miRNA expression profile to characterize the ensemble of both known and novel miRNAs expressed in CRCs, as well as in COCs, by using high-throughput Solexa technology.

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