Browse
Submit Data
Databases
API
Help

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

NULISA: a novel proteomic liquid biopsy platform with attomolar sensitivity and high multiplexing

ABSTRACT: The blood proteome holds great promise for precision medicine but poses daunting challenges due to the low abundance of the majority of plasma proteins and the vast dynamic range across the proteome. We report the development and validation of a novel proteomic analysis technology – NUcleic acid Linked Immuno-Sandwich Assay (NULISA™) – that incorporates a dual capture and release mechanism to suppress the assay background to the minimum, thus drastically improving the signal-to-noise ratio. NULISA improves the sensitivity of the proximity ligation assay by over 10,000-fold to the attomolar level, which is enabled by antibody-conjugated DNA sequences that mediate the purification of immunocomplexes and contain target-and sample-specific barcodes for next-generation sequencing-based, highly multiplexed analysis. To demonstrate its performance and utility, we developed a 200-plex NULISA targeting 124 cytokines and chemokines and 80 other immune response-related proteins that demonstrated superior sensitivity for detecting low-abundance proteins and high concordance with other immunoassays. The ultra-high sensitivity enabled the detection of previously difficult-to-detect but biologically important, low-abundance biomarkers in patients with autoimmune diseases and COVID-19. Fully automated NULISA uniquely addresses longstanding challenges in the proteomic analysis of liquid biopsy samples and makes broad and in-depth proteomic analysis accessible to the general research community and future diagnostic applications.

ORGANISM(S): Homo sapiens

PROVIDER: GSE241717 | GEO | 2023/09/29

REPOSITORIES: GEO

ACCESS DATA

Json Xml

Dataset's files

Source:

			Action	DRS
		Other

Items per page:

1 - 1 of 1

Similar Datasets

Global absolute quantification reveals tight regulation of protein expression in single Xenopus eggs

Project description:Recent developments in genomic sequencing technology have enabled comprehensive transcriptome analyses of single cells. In contrast, single cell proteomics analyses have been restricted to targeted analyses, for example using flow cytometry with GFP fusions or mass cytometry. Here, we performed global absolute protein quantification of single Xenopus laevis eggs using mass spectrometry-based proteomics. We quantified over 5800 proteins, thus representing the largest single cell proteome that has been characterized to date. Absolute protein amounts in single eggs are highly comparable, thus indicating a tight regulation of global protein abundance. Comparison between the single-cell proteome and transcriptome reveal poor expression correlation. Finally, we identified 439 proteins that significantly change in abundance during early embryogenesis. Many of these proteins do not show regulation at the transcript level. Altogether, our data reveal that the transcriptome is a poor indicator of the proteome and that protein levels are tightly controlled in Xenopus leavis eggs. RNA-seq in Xenopus laevis of 5 replicates of both single eggs and single embryos.

2014-08-04 | E-GEOD-56586 | biostudies-arrayexpress

Rapid and in-depth proteomic profiling of small extracellular vesicles for ultralow samples

Project description:The form and function of extracellular vesicles (EV) is defined by their proteome. This knowledge is essential to describe and understand EVs, encompassing their marker proteins, capacity as a signalling platform, and utility as diagnostic tools and therapeutic targets. However, EV are low-abundant entities in the secreteome and biological samples, with disease-specific EV proteins often present in low abundance challenging their detection and quantification due to the inherent challenges in dynamic range using mass spectrometry-based strategies. Combined with lack of protein amplification mechanisms, their proteomic studies require upscaling cell cultures or larger volumes of biofluids. Here, we outline high-sensitivity sample preparation and finely tuned LC gradients for DIA to obtain precise and comprehensive proteome of EVs from ultra-low sample amounts (sub-nanogram).

2023-08-16 | MSV000092668 | MassIVE

Global absolute quantification reveals tight regulation of protein expression in single Xenopus eggs

2014-08-04 | GSE56586 | GEO

Multimodal Omics Elucidation of Metabolic Transition during the Development of Hepatic Organoids

Project description:Hepatic organoids (HO） serve as optimal models simulating liver development, where elucidating the changes in transcriptome and proteome during their development is crucial for understanding the in vitro simulation of liver biological development. Given that HO are embedded in a matrix gel during their maturation stage, extracting metabolites while fully preserving the cellular state poses significant challenges. Through transcriptomics, it is possible to infer alterations in metabolic flux, offering an alternative perspective on metabolic transformation. In this study, we conducted transcriptomic and proteomic sequencing across four developmental stages of HO, analyzed differential genes and proteins, the correlation between transcriptome and proteome, the transcription-translation ratio, and changes in metabolic flux.

2024-12-27 | GSE264422 | GEO

Temperature sensitive Mutant Proteome Profiling: a novel tool for the characterization of the global impacts of missense mutants on the proteome

Project description:RNA-seq experiments measuring global RNA abundance Temperature sensitive (TS) mutants are a tool that have been foundational for the study of many essential life processes. Despite the long-term use of TS mutants, the mechanisms that lead to temperature sensitivity are not fully understood. Furthermore, a high-throughput workflow to characterize biophysical changes occurring in TS mutants is lacking. We developed Temperature sensitive Mutant Proteome Profiling (TeMPP), a novel application of mass spectrometry based thermal proteome profiling (TPP) as a way to measure the effects of missense mutations on protein stability and protein-protein interactions. This study characterized the global changes in mRNA abundance, protein abundance, and protein thermal stability as a result of missense mutants within two subunits of the yeast ubiquitin-proteasome system. Global protein abundance measurements and RNA sequencing data resulted in a large number of possible candidates that could be causing the phenotypic changes observed in the mutant strains. The additional information gained from TeMPP along with complementary proteomic and transcriptomic experiments allows for multiomic intersection analysis that may reveal interesting regulatory categories to pursue in follow-up mechanistic experiments.

2020-01-08 | GSE143236 | GEO

Rapid High-pH Reverse Phase StageTip for Sensitive Small-Scale Membrane Proteomic Profiling

Project description:Membrane proteins are crucial targets for cancer biomarker discovery and drug development. However, in addition to the inherent challenges of hydrophobicity and low abundance, complete membrane proteome coverage of clinical specimen is usually hindered by the requirement of large amount of starting materials. Towards comprehensive membrane proteomic profiling for small amounts of samples (< 10 µg), we developed high-pH reverse phase (Hp-RP) combined with stop-and-go extraction tip (StageTip) technique, as a fast (~ 15 mins.), sensitive, reproducible, high-resolution and multiplexed fractionation method suitable for accurate quantification of the membrane proteome. This approach provided almost 2-fold enhanced detection of peptides within transmembrane helix domains compared with strong anion exchange (SAX) and strong cation exchange (SCX) StageTip techniques. More than 5000 proteins (63% membrane proteins) can be identified in only 10 µg of membrane protein digests, showing the superior sensitivity of the Hp-RP StageTip approach. The Hp-RP StageTip method enabled in-depth membrane proteome profiling of 11 lung cancer cell lines harboring different EGFR mutation status, which resulted in the identification of 3983 annotated membrane proteins. This provides the largest collection of reference peptide spectral data for lung cancer membrane sub-proteome. Finally, relative quantification of membrane proteins between Gefitinib-resistant and -sensitive lung cancer cell lines revealed several upregulated membrane proteins with key roles in lung cancer progression.

2015-11-16 | PXD001548 | Pride

Enhanced Protein Translation Underlies Improved Metabolic and Physical Adaptations to Different Exercise Training Modes in Young and Old Humans

Project description:The molecular transducers of benefits from different exercise modalities remain incompletely defined. Here we report that 12 weeks of high-intensity aerobic interval (HIIT), resistance (RT), and combined exercise training enhanced insulin sensitivity and lean mass, but only HIIT and combined training improved aerobic capacity and skeletal muscle mitochondrial respiration. HIIT revealed a more robust increase in gene transcripts than other exercise modalities, particularly in older adults, although little overlap with corresponding individual protein abundance was noted. HIIT reversed many age-related differences in the proteome, particularly of mitochondrial proteins in concert with increased mitochondrial protein synthesis. Both RT and HIIT enhanced proteins involved in translational machinery irrespective of age. Only small changes of methylation of DNA promoter regions were observed. We provide evidence for predominant exercise regulation at the translational level, enhancing translational capacity and proteome abundance to explain phenotypic gains in muscle mitochondrial function and hypertrophy in all ages.

2017-03-28 | GSE97084 | GEO

Integrated transcriptomic and proteomic analysis of human eccrine sweat glands

Project description:Eccrine sweat gland is an exocrine gland that is involved in the secretion of sweat for control of temperature. Malfunction of the sweat glands can result in disorders such as miliaria, hyperhidrosis and bromhidrosis. In addition, lack of reabsorption of Cl- ions from reabsorptive duct of eccrine sweat gland is a major feature of cystic fibrosis. Understanding the proteome of eccrine sweat glands is important for understanding the physiology of sweat formation. In spite of this, no systematic transcriptome or proteome analysis of eccrine sweat glands has yet been reported. To this end, we isolated eccrine sweat glands by microdissecting them from human skin and performed both RNA-seq and proteome analysis. In total, ~138,000 transcripts and ~6,100 proteins were identified. The proteome data showed the enrichment in protein digestion/absorption and salivary secretion, while the transcriptome data did not show any enrichment for a specific pathway. This study also enabled us to confirm 2 missing proteins. Integrating RNA-seq and proteomic data allowed us to identify 7 peptides from 5 novel genes. Most of the novel proteins were from short open reading frames (sORFs) suggesting that many sORFs still remain to be annotated in the human genome. The peptides mapping to the missing or novel proteins were validated by analyzing synthetic peptides. This study provides the first integrated analysis of the transcriptome and proteome of the human eccrine sweat gland and should become an invaluable resource to biomedical research community for studying sweat glands in physiology and disease.

2018-07-31 | GSE117885 | GEO

Effect of the depletion of ZC3H41 (Tb927.11.1980) on the transcriptome of procyclic Trypanosome brucei

Project description:Depletion of ZC3H41 has minor effects on the transcriptome of procyclic trypanosomes. In general, ZC3H41-regulated transcripts seem to be expressed at low abundance, and almost half of them (43%) are probably non-coding, since the corresponding encoded proteins have not been detected in proteomic surveys . A transcript encoding a trans-sialidase (Tb927.11.11410) was clearly upregulated in ZC3H41-depleted cells .

2023-03-29 | GSE212921 | GEO

Proteogenomics and differential ion mobility enables the exploration of the mutational landscape in colon cancer cells

Project description:The sensitivity and depth of proteomic analyses is limited by isobaric ions and interferences that preclude the identification of low abundance peptides. Extensive sample fractionation is often required to extend proteome coverage when sample amount is not a limitation. Ion mobility devices provide a viable alternate approach to resolve confounding ions, improve peak capacity and mass spectrometry (MS) sensitivity. Here, we report the integration of differential ion mobility with segmented ion fractionation (SIFT) to enhance the comprehensiveness of proteomic analyses. The combination of differential ion mobility and SIFT, where narrow windows of ~ m/z 100 are acquired in turn, is found particularly advantageous in the analysis of protein digests, and typically provided 70% gain in identification compared to conventional single-shot LC-MS/MS. The application of this approach is further demonstrated for the analysis of tryptic digests from different colorectal cancer cell lines where the enhanced sensitivity enabled the identification of single amino acid variants that were correlated with the corresponding transcriptomic datasets.

2023-02-21 | PXD033710 | Pride

OmicsDI is part of the ELIXIR infrastructure

OmicsDI is an Elixir interoperability service. Learn more ›

Tweets

OmicsDI Databases

PRIDE
PeptideAtlas
MassIVE
JPOST Repository
Physiome Model Repository

EGA
EVA
ENA
LINCS
PAXDB
Cell Collective

MetaboLights
Metabolomics Workbench
MetabolomeExpress
GNPS
BioModels
FAIRDOMHub

ArrayExpress
dbGaP
ExpressionAtlas
GEO
NODE

Information

Databases
Help
API
Contact us
Code on GitHub
Terms of use
Submit Data