Self‐made frits for nanoscale columns in proteomics

Maiolica, Alessio; Borsotti, Dario; Rappsilber, Juri

doi:10.1002/pmic.200402010

Cited by 86 publications

(74 citation statements)

References 16 publications

(14 reference statements)

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“…The LC was operated in the two-column mode. The homemade fused silica column equipped with a glass fiber frit was packed with Reprosil-Pur 120 C18-AQ 3 lm resin and connected to the analytical column via an ultra high-pressure liquid chromatography (UHPLC) union (50). The analytical column was a fused silica capillary (75 lm · 25 cm) with integrated PicoFrit emitter packed in-house with Reprosil-Pur 120 C18-AQ 3 lm resin.…”

Section: Liquid Chromatography-ms/msmentioning

confidence: 99%

Mesenchymal Stem Cell Therapy Protects Lungs from Radiation-Induced Endothelial Cell Loss by Restoring Superoxide Dismutase 1 Expression

Klein

Steens

Wiesemann

et al. 2017

Antioxidants & Redox Signaling

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Aims: Radiation-induced normal tissue toxicity is closely linked to endothelial cell (EC) damage and dysfunction (acute effects). However, the underlying mechanisms of radiation-induced adverse late effects with respect to the vascular compartment remain elusive, and no causative radioprotective treatment is available to date. Results: The importance of injury to EC for radiation-induced late toxicity in lungs after whole thorax irradiation (WTI) was investigated using a mouse model of radiation-induced pneumopathy. We show that WTI induces EC loss as long-term complication, which is accompanied by the development of fibrosis. Adoptive transfer of mesenchymal stem cells (MSCs) either derived from bone marrow or aorta (vascular wall-resident MSCs) in the early phase after irradiation limited the radiation-induced EC loss and fibrosis progression. Furthermore, MSC-derived culture supernatants rescued the radiation-induced reduction in viability and longterm survival of cultured lung EC. We further identified the antioxidant enzyme superoxide dismutase 1 (SOD1) as a MSC-secreted factor. Importantly, MSC treatment restored the radiation-induced reduction of SOD1 levels after WTI. A similar protective effect was achieved by using the SOD-mimetic EUK134, suggesting that MSCderived SOD1 is involved in the protective action of MSC, presumably through paracrine signaling. Innovation: In this study, we explored the therapeutic potential of MSC therapy to prevent radiation-induced EC loss (late effect) and identified the protective mechanisms of MSC action. Conclusions: Adoptive transfer of MSCs early after irradiation counteracts radiation-induced vascular damage and EC loss as late adverse effects. The high activity of vascular wall-derived MSCs for radioprotection may be due to their tissue-specific action. Antioxid. Redox Signal. 26, 563-582.

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Section: Liquid Chromatography-ms/msmentioning

confidence: 99%

Mesenchymal Stem Cell Therapy Protects Lungs from Radiation-Induced Endothelial Cell Loss by Restoring Superoxide Dismutase 1 Expression

Klein

Steens

Wiesemann

et al. 2017

Antioxidants & Redox Signaling

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“…The column was then clipped to the desired final length. The permanent inlet frit was then prepared using a procedure described by Maiolica et al [33] The column inlet was gently pressed on a glass microfiber filter (Reeve Angel; Clifton, NJ) that was previously wetted with 1:1 (v:v) ratio of potassium silicate (Kasil) and formamide. The Kasil was used as received from PQ Corporation (Valley Forge, IA) and formamide was from Sigma-Aldrich (St. Louis, MO).…”

Section: Column Packingmentioning

confidence: 99%

1.1 μm Superficially porous particles for liquid chromatography

Blue

Jorgenson

2015

Journal of Chromatography A

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“…Electrospray emitters were custom made using 150 m outer diameter ϫ 20 m inner diameter chemically etched fused silica capillaries, as previously described (19). Reversed-phase capillary columns were prepared by slurry packing 3-m Jupiter C18 bonded particles (Phenomenex, Torrance, CA) into a 75 m ϫ 65 cm fused silica capillary (Polymicro Technologies, Phoenix, AZ) using 0.5 cm sol-gel plugs for particle retention (20). Mobile phases consisted of (a) 0.1% formic acid in water and (b) 0.1% formic acid in acetonitrile and were degassed on-line using a Degasys Model DG-2410 vacuum degasser (Dionex, Germany); the HPLC system was equilibrated at 10,000 psi with 100% mobile phase (a) for initial starting conditions.…”

Section: Methodsmentioning

confidence: 99%

Bayesian Proteoform Modeling Improves Protein Quantification of Global Proteomic Measurements

Webb-Robertson

Matzke

Datta

et al. 2014

Molecular & Cellular Proteomics

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As the capability of mass spectrometry-based proteomics has matured, tens of thousands of peptides can be measured simultaneously, which has the benefit of offering a systems view of protein expression. However, a major challenge is that, with an increase in throughput, protein quantification estimation from the native measured peptides has become a computational task. A limitation to existing computationally driven protein quantification methods is that most ignore protein variation, such as alternate splicing of the RNA transcript and post-translational modifications or other possible proteoforms, which will affect a significant fraction of the proteome. The consequence of this assumption is that statistical inference at the protein level, and conse- The application of MS-based proteomics has resulted in large-scale studies in which the set of measured, and subsequently identified, peptides is often used to estimate protein abundance. In particular, label-free MS-based proteomics is highly effective for identification of peptides and measurement of relative peptide abundances (1, 2), but it does not directly yield protein quantities. The importance of accurate protein quantification cannot be understated; it is the essential component of identifying biomarkers of disease or defining the relationship between gene regulations, protein interactions, and signaling networks in a cellular system (3, 4). The major challenge is that protein abundance depends not only on transcription rates of the gene but also on additional control mechanisms, such as mRNA stability, translational regulation, and protein degradation. Moreover, the functional activity of proteins can be altered through a variety of posttranslational modifications or proteolytic processing and alternative splicing, events which selectively alter the abundance of some selected peptides while leaving others unchanged (4). This complexity of the proteome, in addition to issues associated with the measurement and identification

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Self‐made frits for nanoscale columns in proteomics

Cited by 86 publications

References 16 publications

Mesenchymal Stem Cell Therapy Protects Lungs from Radiation-Induced Endothelial Cell Loss by Restoring Superoxide Dismutase 1 Expression

Mesenchymal Stem Cell Therapy Protects Lungs from Radiation-Induced Endothelial Cell Loss by Restoring Superoxide Dismutase 1 Expression

1.1 μm Superficially porous particles for liquid chromatography

Bayesian Proteoform Modeling Improves Protein Quantification of Global Proteomic Measurements

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