Development of High-Throughput Chemical Extraction Techniques and Quantitative HPLC-MS/MS (SRM) Assays for Clinically Relevant Plasma Proteins

2010 ◽  
Vol 9 (1) ◽  
pp. 333-340 ◽  
Author(s):  
Chris Barton ◽  
Richard G. Kay ◽  
Wolfgang Gentzer ◽  
Frank Vitzthum ◽  
Steve Pleasance
Keyword(s):  
High Throughput ◽  
Plasma Proteins ◽  
Chemical Extraction ◽  
Extraction Techniques ◽  
Quantitative Hplc

The application of nuclear magnetic resonance to the study of cellular physiology

1984 ◽  
Vol 246 (1) ◽  
pp. C10-C19 ◽  
Author(s):  
R. S. Balaban
Keyword(s):  
Nuclear Magnetic Resonance ◽  
Magnetic Resonance ◽  
Inorganic Compounds ◽  
Chemical Extraction ◽  
Kinetic Properties ◽  
Extraction Techniques ◽  
Isolated Cells ◽  
Cellular Physiology ◽  
Nuclear Magnetic

Nuclear magnetic resonance (NMR) is a form of absorption spectroscopy that can noninvasively monitor the intracellular concentration and several kinetic properties of numerous organic and inorganic compounds. Utilizing these characteristics, investigators have demonstrated that NMR is a useful tool in the study of cellular physiology. In this review, the techniques for using NMR to study isolated cells are outlined with suggestions for the determination of cellular viability within the NMR spectrometer. Whenever feasible, cell preparations that are continuously perfused are preferred, because they can be constantly fed and controlled. Results of various NMR experiments on isolated cells using several nuclides are reviewed to highlight the type of information NMR can provide about cellular physiology. Several important differences between NMR and chemical extraction data are noted. The reason for these differences is probably related to the chemical extraction techniques determining the total amount of a compound within the cell in comparison to NMR, which is somewhat more specific, theoretically, detecting only the free species within the cytosol.

Partitioning and Leaching Behavior of Actinides and Rare Earth Elements in a Zirconolite-bearing Hydrothermal Vein System

2006 ◽  
Vol 985 ◽  
Author(s):  
Timothy E Payne ◽  
Reto Giere ◽  
Kaye P Hart ◽  
Gregory R Lumpkin ◽  
Peter J McGlinn
Keyword(s):  
Rare Earth ◽  
Rare Earth Elements ◽  
Major Elements ◽  
Chemical Extraction ◽  
Extraction Techniques ◽  
Leaching Behavior ◽  
Bulk Chemical Composition ◽  
Major Vein ◽  
Bulk Chemical ◽  
And Behavior

AbstractChemical extraction techniques and scanning electron microscopy were used to study the distribution and behavior of actinides and rare earth elements (REE) in hydrothermal veins at Adamello, (Italy). The six samples discussed in this paper were from the phlogopite zone, which is one of the major vein zones. The samples were similar in their bulk chemical composition, mineralogy, and leaching behavior of major elements (determined by extraction with 9M HCl). However, there were major differences in the extractability of REE and actinides. The most significant influence on the leaching characteristics appears to be the amounts of U, Th and REE incorporated in resistant host phases. Uranium and Th are very highly enriched in zirconolite grains. Actinides were more readily leached from samples with a higher content of U and Th, relative to the amount of zirconolite. The results show that REE and actinides present in chemically resistant minerals can be retained under aggressive leaching conditions.

scholarly journals iTRAQ-based high-throughput proteomics analysis reveals alterations of plasma proteins in patients infected with human bocavirus

PLoS ONE ◽  
2019 ◽  
Vol 14 (11) ◽  
pp. e0225261
Author(s):  
Junmei Bian ◽  
Min Liang ◽  
Shuxian Ding ◽  
Liyan Wang ◽  
Wenchang Ni ◽  
...  
Keyword(s):  
High Throughput ◽  
Plasma Proteins ◽  
Human Bocavirus ◽  
Proteomics Analysis

Evaluation of PQ500, a 500-plasma protein blood panel in NSCLC subjects using high-throughput MRM mass spectrometry.

2019 ◽  
Vol 37 (8_suppl) ◽  
pp. 110-110
Author(s):  
Nicholas Dupuis ◽  
Sebastian Muller ◽  
Tobias Treiber ◽  
Claudia Escher
Keyword(s):  
High Throughput ◽  
Plasma Proteins ◽  
Capillary Flow ◽  
Stage Iv ◽  
Absolute Quantification ◽  
Response To Therapy ◽  
Statistical Testing ◽  
Triple Quadrupole Mass Spectrometer ◽  
Amyloid A ◽  
Key Species

110 Background: Monitoring of circulating proteins in cancer subjects can provide insight into the host response to the tumor. Recent studies have demonstrated the utility of circulating proteins for early detection, differential diagnosis, and predicting response to therapy. Although the utility is clear, standardization of measurement methods across a large number of proteins from plasma has remained challenging. Here, we evaluate the detection of proteins from a > 500-protein plasma panel assembled from stable-isotope standard (SIS) peptides and measured in a high-throughput MRM workflow. Methods: Plasma samples from subjects with Stage III-IV non-small cell lung cancer (n = 15), Stage IV pancreatic cancer (n = 6) and healthy donors (n = 15) were prepared for analysis. Prior to analysis, a panel of SIS peptides covering 582 plasma proteins (804 peptides), was spiked into each sample enabling absolute quantification of detectable peptide species. All samples were analyzed using a capillary flow liquid chromatography (5 µl/min) with a one-hour gradient coupled to a Thermo Scientific TSQ Altis triple quadrupole mass spectrometer. Protein data was extracted and quantified for statistical analysis using SpectroDive (Biognosys). Results: Across all samples, 607 peptides representing 411 proteins were quantified at least once, with an average of 300 proteins quantified per sample. From triplicate analyses, the median CV across all proteins was 5.3%. Statistical testing identified 42 significantly regulated proteins (36 up- and 6 down-regulated; q-value > 0.05 and log2 fold change > 0.58). Key species that were strongly dysregulated in cancer samples included: c-reactive protein (CRP), serum amyloid a (SAA1/SAA2), complement C9, S100A8/S100A9, and leucine rich glycoprotein 1 (LRG1). Conclusions: PQ500 is a comprehensive blood-focused multiplexed biomarker panel that enables absolute quantification of plasma proteins. The dysregulated proteins identified in this study highlight that the PQ500 can monitor the host immune response through multiple pathways. Accurate monitoring these proteins offers the possibility to define molecular based markers with clinical and diagnostic utility.

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