Dataset Information

MALDI-MS Profiling of Intact Blood Proteins without the Need for Freezing - Dried Serum Spots as Future Clinical Tools for Patient Screening

ABSTRACT: To open new ways for MALDI-MS-based patient screening, blood serum is the most preferred specimen because of its richness in patho-physiological information and due to ease of collection. To overcome deleterious freeze / thaw cycles and to reduce high costs for shipping and storage, we sought to develop a procedure which enables MALDI-MS protein profiling of blood serum proteins without the need for serum freezing. MALDI mass spectra from either (p)FFS or (p)DSS preparations showed on average 25 strong ion signals (mass range m/z 6000 to m/z 10,000) from intact serum proteins (apolipoproteins, complement proteins, transthyretin and hemoglobin) and from proteolytic processing products. Semi-quantitative analysis of three ion pairs: m/z 6433 and 6631, m/z 8205 and 8916, as well as m/z 9275 and 9422 indicated that the mass spectra from either (p)FFS or (p)DSS contained the same protein composition information. A workflow that avoids the conventional cold-chain and yet enables to investigate intact serum proteins and/or serum proteolysis products by MALDI-MS profiling was developed. The here described protocol tremendously broadens clinical application of MALDI-MS and simultaneously allows to reduce the costs for storage and shipping of serum samples.

INSTRUMENT(S): Bruker Daltonics flex series

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Blood

SUBMITTER: Michael Kreutzer

LAB HEAD: Michael O. Glocker

PROVIDER: PXD024669 | Pride | 2021-05-20

REPOSITORIES: Pride

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Profiling of intact blood proteins by matrix-assisted laser desorption/ionization mass spectrometry without the need for freezing - Dried serum spots as future clinical tools for patient screening.

Okai Charles A CA Wölter Manja M Russ Manuela M Koy Cornelia C Petre Brindusa A BA Rath Werner W Pecks Ulrich U Glocker Michael O MO

Rapid communications in mass spectrometry : RCM 20210701 14

<h4>Rationale</h4>To open up new ways for matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS)-based patient screening, blood serum is the most preferred specimen because of its richness in patho-physiological information and due to ease of collection. To overcome deleterious freeze/thaw cycles and to reduce high costs for shipping and storage, we sought to develop a procedure which enables MALDI-MS protein profiling of blood serum proteins without the need for serum freezing ...[more]

PMID: 33955049

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Project description:Background: MicroRNAs (miRNAs) acting as negative regulators of gene expression are differentially expressed in intestinal tissues of patients with inflammatory bowel disease (IBD). Assessing the functional role of miRNAs in murine models of colitis facilitates elucidating the role of specific miRNAs in human IBD. The aim of this study was to determine the miRNA signature of murine models of colitis and to assess the influence of miR-21 on intestinal inflammation. Methods: miRNAs expression was accessed by microarray for acute and chronic murine model of colitis induced by DSS or TNBS. miR-21-deficient mouse and littermates controls were assessed in the standard DSS, TNBS and CD4+ T cell transfer models of colitis. RNAs of mouse colon and CD4+CD45RBHigh cells were analyzed by miRNA and mRNA microarray, and quantitative RT-PCR. Th1 polarization was accessed by flow-cytometry and ELISA. Results: Alterations of in miRNAs expression were identified for acute and chronic DSS colitis and TNBS colitis, receptively. The Expression of miRs-21, -142-3p and -223 was were distinct between DSS and TNBS models while overlap of numerous miRNAs was seen. Importantly, miRs-19b, -192 and -215, that are decreased in IBD, were significantly decreased in all 4 models of colitis. miR-21, which is increased in IBD, was increased in TNBS colitis but not the DSS colitis models. Further assessment of the miR-21-deficient 1-/- mice revealed that the deletion of miR-21 results in the exacerbation of both the TNBS and T cell-transfer models of colitis. Conclusions: miRNAs are differentially expressed in both human IBD and murine colitis, with overlap of several IBD-associated miRNAs. The demonstration that miR-21 deletion exacerbated CD4+ T cell-mediated models of colitis provides further evidence that miRNAs play significant roles in the pathogenesis of IBD. miRNAs expression was accesed for acute and chronic murine model of colitis induced by DSS or TNBS.Total of 20 samples with duplicates were analyed in this study.

			Action	DRS
	DSS.FID.zip	Other
	F416453.dat	Other
	F416453.mgf	Mgf
	FFS.FID.zip	Other
	Okai_patient_numbers_and_target_positions.txt	Txt