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    • "Although single-marker diagnostic indicators provide the most straightforward assessment of disease, they are rarely able to provide a reliable and complete biological snapshot necessary to characterize diseases (Pepe et al., 2001). Multiplexed approaches that test for many markers simultaneously have been shown to be more sensitive and accurate in detecting and staging complex diseases like cancer, even surpassing the performance of today's current practice (Cao et al., 2011; Shimizu et al., 2012). "
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    ABSTRACT: The full utility of miRNA profiling has yet to be realized largely due to the limited throughput of existing validation technologies, errors introduced during sample purification and insufficient methods used for analytical normalization and quality control. We introduce Abcam's Firefly™ particle technology with assays that enable researchers to rapidly and cost-effectively evaluate miRNA biomarker signatures on standard flow cytometers. In order to accommodate studies in solid tissues and circulating biofluids, we developed the Multiplex Cellular miRNA Assay and Multiplex Circulating miRNA Assay, respectively. We demonstrate that these assays provide miRNA profiles that correlate well with currently available miRNA profiling technology and show excellent reproducibility even with diminishing RNA input. We have integrated bioinformatics tools into our assays to effectively interpret experimental data, help identify candidate signatures and produce publication-quality figures.
    Full-text · Technical Report · Oct 2015
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    • "Therefore, the composition of a given proteome can be altered depending on the biological state of the organism [2]. Thus, protein identification as well as relative or absolute quantification of a given protein in different biological conditions, i.e. healthy and diseased states [3], and the identification of post-translational changes are the basis for understanding the fundamental processes that occur in living organisms [4]. The development of new analytical–chemical strategies for protein analysis is important for progress in proteomics [5]. "
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    ABSTRACT: Labeling of peptides and proteins using chelators binding metal ions has become a novel approach for quantitative proteomics in recent years. The aim of this work was the optimization of a new method for peptide derivatization with lanthanide labels (holmium, lutetium, and thulium) followed by nano high performance liquid chromatography (nano-HPLC) separation with UV detection. Matrix-assisted laser desorption ionization–mass spectrometry (MALDI MS) was used to confirm the derivatization and to identify the derivatized peptides. Peptides were labeled with the three different lanthanide metals using a bifunctional DOTA-based (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid) reagent. The results demonstrate that the derivatization reaction using the chelating reagent DOTA-NHS-ester was effective for single peptides and peptide mixtures. Furthermore, an efficient pre-cleaning method was applied by nano-HPLC using a C-18 trap column for elimination of the excess of labeling reagent. The application of the optimized method to label peptides in a Cytochrome C digest delivered comparable results to those obtained with model peptides.
    Full-text · Article · Jan 2015 · Microchemical Journal
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    • "ideal biomarker should have the following aspects in order to be able to predict or diagnose a specific dis - ease or condition ( Pepe et al . , 2001 ) : 1 ) simple and safe assays ; 2 ) provide guidance to ease decision - making ; 3 ) establish correlation between an assessment result and a clinical condition ( Fletcher et al . , 2012 ) in a sen - sitive ( a positive outcome when the disease is present ) and a specific ( a negative outcome when the disease is absent ) way . As with "
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    ABSTRACT: Glycans are chains of carbohydrates attached to proteins (glycoproteins and proteoglycans) or lipids (glycolipids). Glycosylation is a post-translational modification and glycans have a wide range of functions in the human body including involvement in oncological diseases. Change in a glycan structure can not only indicate the presence of a pathological process but, more importantly, in some cases also its stage. Thus, a glycan analysis has the potential to be an effective and reliable tool in cancer diagnostics. Lectins are proteins responsible for natural biorecognition of glycans; even carbohydrate moieties still attached to proteins or whole cells can be recognised by lectins, which makes them an ideal candidate for designing label-free biosensors for glycan analysis. This review seeks to summarise evidence that the glycoprofiling of biomarkers by lectin-based biosensors can be of significant help in detecting prostate cancer.
    Full-text · Article · Jan 2015 · Chemical Papers
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