Browse
Submit Data
Databases
API
Help

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

Compatibility of distinct label-free proteomic workflows in absolute quantification of proteins linked to the oocyte quality in human follicular fluid (the Triple-Quad TOF workflow)

ABSTRACT: We present a comprehensive analysis of human follicular fluid proteome by comparing several strategies of sample preparation, mass spectrometry analysis, and data processing, resulting in the identification of 1638 proteins. We devised two separate label-free quantitative workflows, termed after the utilized mass spectrometer: Triple Quad-TOF and Quad-Orbitrap, and analyzed their performance in absolute protein quantification assisted by Total Protein Approach. In a pilot study of twenty samples linked to diverse oocyte quality status from four donors, 215 and 455 proteins were quantified by the Triple Quad-TOF and Quad-Orbitrap workflows, respectively. The concentration values obtained from both workflows correlated to a significant degree. We found reasonable agreement of both workflows in protein fold changes between tested groups, resulting in unified lists of 21 and 24 proteins, linked to oocyte maturity and blastocyst development, respectively. The Quad-Orbitrap workflow was best suited for an in-depth analysis, especially of low abundant proteome, whereas the Triple Quad-TOF workflow allowed a more robust approach with a greater potential to increase in effectiveness with the growing number of analyzed samples.

INSTRUMENT(S): TripleTOF 5600

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Follicular Fluid

SUBMITTER: Aleksandra Bogucka

LAB HEAD: Stanisław Oldziej

PROVIDER: PXD024223 | Pride | 2021-07-23

REPOSITORIES: Pride

ACCESS DATA

Json Xml

Dataset's files

Source:

			Action	DRS
	3TOF_HFF_IS.xlsx	Xlsx
	3TOF_HFF_LTC.xlsx	Xlsx
	3TOF_HFF_T.xlsx	Xlsx
	3TOF_HMW_T.xlsx	Xlsx
	Clinical.xlsx	Xlsx

Items per page:

1 - 5 of 20

Similar Datasets

Compatibility of distinct label-free proteomic workflows in absolute quantification of proteins linked to the oocyte quality in human follicular fluid (the Quad-Orbitrap workflow)

Project description:We present a comprehensive analysis of human follicular fluid proteome by comparing several strategies of sample preparation, mass spectrometry analysis, and data processing, resulting in the identification of 1638 proteins. We devised two separate label-free quantitative workflows, termed after the utilized mass spectrometer: Triple Quad-TOF and Quad-Orbitrap, and analyzed their performance in absolute protein quantification assisted by Total Protein Approach. In a pilot study of twenty samples linked to diverse oocyte quality status from four donors, 215 and 455 proteins were quantified by the Triple Quad-TOF and Quad-Orbitrap workflows, respectively. The concentration values obtained from both workflows correlated to a significant degree. We found reasonable agreement of both workflows in protein fold changes between tested groups, resulting in unified lists of 21 and 24 proteins, linked to oocyte maturity and blastocyst development, respectively. The Quad-Orbitrap workflow was best suited for an in-depth analysis, especially of low abundant proteome, whereas the Triple Quad-TOF workflow allowed a more robust approach with a greater potential to increase in effectiveness with the growing number of analyzed samples.

2021-07-23 | PXD024347 | Pride

Proteomic profiling of NCI60 cell lines from Cancer Cell Line Encyclopedia

Project description:This ArrayExpress record contains meta-data and results of quantitative analysis of cell lines from the NCI-60 panel using pressure cycling technology (PCT) and SWATH-mass spectrometry. Each cell line was analyzed in duplicate. Raw data files are available at the EMBL-EBI protemics data archive (PRIDE) at accession PXD003539 (http://www.ebi.ac.uk/pride/archive/projects/PXD003539). Since the record here does not include the raw data files and hence there is no need to explicitly link individual replicate to a raw file, each sample is only listed once in the ArrayExpress samples table for clarity.

2017-01-27 | E-PROT-2 | biostudies-arrayexpress

Proteome and peptidome of human follicular fluid

Project description:Human follicular fluid (hFF) is a natural environment of oocyte maturation, and some components of hFF could be used to judge oocyte capability for fertilization and further development. In our pilot small scale study three samples from four donors (12 samples in total) were analyzed to determine which hFF proteins/peptides could be used to differentiate individual oocytes and which are patient specific. Ultrafiltration was used to fractionate hFF to High Molecular Weight (HMW) – proteome (>10kDa) and Low Molecular Weight (LMW) – peptidome (<10 kDa) fractions. HMW and LMW compositions were analyzed using LC-MS in SWATH data acquisition and processing methodology. In total we were able to identify 158 proteins, form which 59 were never reported before as hFF components. 55 (45 Never reported before) proteins were found by analyzing LMW fraction, 67 (14 never reported before) were found by analyzing HMW fraction, and 36 were identified in both fractions of hFF. We were able to perform quantitative analysis for 72 proteins from HMW fraction of hFF. We found that concentrations of 18 proteins varied substantially among hFF samples from single donors and those proteins are promising targets to identify biomarkers useful in oocyte quality assessment.

2017-07-04 | PXD006550 | Pride

Label free SWATH proteomics reveals sex-biased proteins potentially associated with sex expression and modification in dioecious C. grandis

Project description:Dioecy is an important sexual system wherein, male and female flowers are borne on separate unisexual plants. Knowledge of sex-related differences can enhance our understanding in molecular and developmental processes leading to unisexual flower development. Coccinia grandis is a dioecious species belonging to Cucurbitaceae, a family well-known for diverse sexual systems. Male and female plants of C. grandis have 22A+XY and 22A+XX chromosomes respectively. Previously, we have reported a gynomonoecious form (GyM) (22A+XX) of C. grandis bearing morphologically hermaphrodite flowers (GyM-H) and female flowers (GyM-F). Also, we showed that foliar spray of silver nitrate on female C. grandis plant induces development of morphologically hermaphrodite buds (Ag-H) despite the absence of Y chromosome. To identify sex-related differences, total protein from the flower buds of male, female, GyM-H and Ag-H of C. grandis at early and middle stages of development were analysed by a powerful label-free proteomics approach on ABSCIEX Triple TOF 5600 platform.

2019-07-29 | PXD011064 | Pride

Red blood cells protein profile is modified in breast cancer patients

Project description:We characterized the proteomic composition of RBCs from 53 breast cancer patients (stages I-III and IV), compared with 33 healthy donors. For that, we performed two different proteomic analyses (LC-MS/MS in DDA mode-shotgun- and a SWATH-MS) using the Triple TOF 6600 technology.

2023-01-17 | PXD030936 | Pride

Lon protease is important for growth under stressful conditions and pathogenicity of the phytopathogen, bacterium Dickeya solani.

Project description:The Lon protein is a protease implicated in virulence of many pathogenic bacteria, including some plant pathogens. However, little is known about the role of Lon in bacteria from genus Dickeya. This group of bacteria include important potato pathogens, with the most aggressive species, D. solani. To determine the importance of Lon for pathogenicity and response to stress conditions of bacteria, we constructed a D. solani Δlon strain. The mutant bacteria showed increased sensitivity to certain stress conditions, in particular osmotic and high-temperature stresses. Furthermore, qPCR analysis showed an increased expression of the lon gene in D. solani under these conditions. The deletion of the lon gene resulted in decreased motility, lower activity of secreted pectinolytic enzymes and finally delayed onset of blackleg symptoms in the potato plants. In the Δlon cells, the altered levels of several proteins, including virulence factors and proteins associated with virulence, were detected by means of MS-SWATCH analysis. These included components of the type III secretion system and proteins involved in bacterial motility. Our results indicate that Lon protease is important for D. solani to withstand stressful conditions and effectively invade the potato plant.

2021-09-09 | PXD018297 | Pride

Comparison of fractionation proteomics for local SWATH library building

Project description:For data-independent acquisition by means of sequential window acquisition of all theoretical fragment ion spectra (SWATH), a reference library of data-dependent acquisition (DDA) runs is typically used to correlate the quantitative data from the fragment ion spectra with peptide identifications. The quality and coverage of such a reference library is therefore essential when processing SWATH data. In general, library sizes can be increased by reducing the impact of DDA precursor selection with replicate runs or fractionation. However, these strategies can affect the match between the library and SWATH measurement, and thus larger library sizes do not necessarily correspond to improved SWATH quantification. Here, three fractionation strategies to increase local library size were compared to standard library building using replicate DDA injection: protein SDS-PAGE fractionation, peptide high-pH RP-HPLC fractionation and MS-acquisition gas phase fractionation. The impact of these libraries on SWATH performance was evaluated in terms of the number of extracted peptides and proteins, the match quality of the peptides and the extraction reproducibility of the transitions. These analyses were conducted using the hydrophilic proteome of differentiating human embryonic stem cells.

2017-07-04 | PXD006190 | Pride

Multipronged quantitative proteomics reveals serum proteome alterations in breast cancer intrinsic subtypes

Project description:Being molecularly heterogeneous, breast cancer tends to be a complicated oncological disease with high incidence rates throughout the world. The primary aim of this study was to identify the set of serum proteins with discriminatory capabilities towards the four major subtypes of breast cancer. We employed multipronged quantitative proteomic approaches like 2D-DIGE, iTRAQ and SWATH-MS and identified 307 differentially regulated proteins. Luminal A subtype consisted of 24, Luminal B subtype 38, HER2 Enriched subtype 17 and Triple negative breast cancer subtype 10 differentially regulated subtype specific proteins. These specific proteins were further subjected to bioinformatic analysis viz. PANTHER, DAVID and LENS which revealed the involvement in platelet degranulation, fibrinolysis, lipid metabolism, immune response, complement activation, blood coagulation, immune cell activation, glycolysis, amino acid biosynthesis and cancer signaling pathways in the subtypes of the breast cancer. The significant discrimination efficiency of the models generated through multivariate statistical analysis was decent to distinguish each of the four subtypes from controls. Further, some of the statistically significant differentially regulated proteins were verified and validated by immunoblotting and mass spectrometry based selected reaction monitoring (SRM) approach. Our Multipronged proteomics approaches revealed panel of serum proteins specifically altered for individual subtypes of breast cancer.

2017-05-15 | PXD006441 | Pride

Quantitative analysis of rtPA-associated ICH protein serum and plasma biomarkers in acute ischemic stroke

Project description:Acute stroke, with ischemic stroke comprising 80% of all cerebrovascular incidents, has been recognized as one of the core problems in clinical medicine in need of prevention and treatment. Intravenous rtPA is the mainstay and the highest class evidence based method of acute ischemic stroke treatment, and is currently recommended 0-4.5 hours after stroke onset. In most patients decision on i.v. rtPA administration is striaghtforward, however the biggest concern is the symptomatic intracranial haemorrhage (sICH), which occurs in 3-7% of all treated patients, and is associated with worse 90-day functional outcome and higher disability than in those untreated. Finding a method of the powerful (highly specific and selective) identification of patients at highest risk of sICH, in order to increase the percentage of stroke patients safely treated with rtPA, is one of the most important challenges in stroke research. To address this problem we designed a major and complex project to identify blood, neuroimaging, and clinical biomarkers combined for prospective assessment of the risk of intracranial hemorrhage (ICH) after thrombolytic treatment of acute ischemic stroke. Herein, we reveal our general methodological approach with shortlisting of blood protein candidates selected with Sequential Window Acquisition of All Theoretical Mass Spectra (SWATH-MS) that in the future might increase sensitivity and selectivity of the rtPA-associated sICH risk calculations.

2021-11-25 | PXD021713 | Pride

Proteomic analysis of filaggrin insufficient keratinocytes and secreted exosomes.

Project description:Filaggrin (FLG) is a key structural protein expressed in epidermal keratinocytes. Mutations in the filaggrin gene (FLG) are strongly linked to atopic dermatitis (AD) and allergic inflammation occurring later in life at distant body locations in AD patients. Keratinocytes secrete exosomes, small, lipid-rich membrane vesicles to communicate with distant cells, including within the immune system. The FLG gene was silenced in the HaCaT keratinocyte cell line by shRNA delivered using a lentiviral vector. HaCaT cells transduced with scrambled shRNA were used as a control.

2022-08-12 | PXD026859 | Pride