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      ELISA in the multiplex era: Potentials and pitfalls

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          Abstract

          Multiplex immunoassays confer several advantages over widely adopted singleplex immunoassays including increased efficiency at a reduced expense, greater output per sample volume ratios and higher throughput predicating more resolute, detailed diagnostics and facilitating personalised medicine. Nonetheless, to date, relatively few protein multiplex immunoassays have been validated for in vitro diagnostics in clinical/point‐of‐care settings. This review article will outline the challenges, which must be ameliorated prior to the widespread integration of multiplex immunoassays in clinical settings: (i) biomarker validation; (ii) standardisation of immunoassay design and quality control (calibration and quantification); (iii) availability, stability, specificity and cross‐reactivity of reagents; (iv) assay automation and the use of validated algorithms for transformation of raw data into diagnostic results. A compendium of multiplex immunoassays applicable to in vitro diagnostics and a summary of the diagnostic products currently available commercially are included, along with an analysis of the relative states of development for each format (namely planar slide based, suspension and planar/microtitre plate based) with respect to the aforementioned issues.

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          Most cited references81

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          Coactivation of receptor tyrosine kinases affects the response of tumor cells to targeted therapies.

          Targeted therapies that inhibit receptor tyrosine kinases (RTKs) and the downstream phosphatidylinositol 3-kinase (PI3K) signaling pathway have shown promising anticancer activity, but their efficacy in the brain tumor glioblastoma multiforme (GBM) and other solid tumors has been modest. We hypothesized that multiple RTKs are coactivated in these tumors and that redundant inputs drive and maintain downstream signaling, thereby limiting the efficacy of therapies targeting single RTKs. Tumor cell lines, xenotransplants, and primary tumors indeed show multiple concomitantly activated RTKs. Combinations of RTK inhibitors and/or RNA interference, but not single agents, decreased signaling, cell survival, and anchorage-independent growth even in glioma cells deficient in PTEN, a frequently inactivated inhibitor of PI3K. Thus, effective GBM therapy may require combined regimens targeting multiple RTKs.
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            Urinary and serum biomarkers for the diagnosis of acute kidney injury: an in-depth review of the literature.

            Acute kidney injury (AKI) remains associated with high morbidity and mortality, despite progress in medical care. Although the RIFLE (Risk, Injury, Failure, Loss, End-Stage Kidney Disease) and AKIN (Acute Kidney Injury Network) criteria, based on serum creatinine and urine output, were a step forward in diagnosing AKI, a reliable tool to differentiate between true parenchymal and pre-renal azotaemia in clinical practice is still lacking. In the last decade, many papers on the use of new urinary and serum biomarkers for the diagnosis and prognostication of AKI have been published. Thus, the question arises which biomarker is a reliable differential diagnostic tool under which circumstances. We searched Medline from inception to April 2012 using medical subject heading and text words for AKI and biomarkers [neutrophil gelatinase-associated lipocalin (NGAL), kidney injury molecule-1 (KIM-1), Cystatin C, interleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-18 (IL-18), N-acetyl-glucosaminidase (NAG), glutathione transferases (GST) and liver fatty acid binding protein (LFABP)] to identify relevant papers in five different settings (paediatrics, cardiac surgery, emergency department, critically ill and contrast-induced nephropathy). We included 87 relevant papers, reporting on 74 studies. Depending upon the setting, 7-27 different definitions of AKI were used. Reported diagnostic performance of the different biomarkers was variable from poor to excellent, and no consistent generalizable conclusions can be drawn on their diagnostic value. Early diagnosing of AKI in clinical conditions by using new serum and urinary biomarkers remains cumbersome, especially in those settings where timing and aetiology of AKI are not well defined. Putting too much emphasis on markers that have not convincingly proven reliability might lead to incorrect interpretation of clinical trials. Further research in this field is warranted before biomarkers can be introduced in clinical practice.
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              Update on the biomarker core of the Alzheimer's Disease Neuroimaging Initiative subjects.

              Here, we review progress by the Penn Biomarker Core in the Alzheimer's Disease Neuroimaging Initiative (ADNI) toward developing a pathological cerebrospinal fluid (CSF) and plasma biomarker signature for mild Alzheimer's disease (AD) as well as a biomarker profile that predicts conversion of mild cognitive impairment (MCI) and/or normal control subjects to AD. The Penn Biomarker Core also collaborated with other ADNI Cores to integrate data across ADNI to temporally order changes in clinical measures, imaging data, and chemical biomarkers that serve as mileposts and predictors of the conversion of normal control to MCI as well as MCI to AD, and the progression of AD. Initial CSF studies by the ADNI Biomarker Core revealed a pathological CSF biomarker signature of AD defined by the combination of Abeta1-42 and total tau (T-tau) that effectively delineates mild AD in the large multisite prospective clinical investigation conducted in ADNI. This signature appears to predict conversion from MCI to AD. Data fusion efforts across ADNI Cores generated a model for the temporal ordering of AD biomarkers which suggests that Abeta amyloid biomarkers become abnormal first, followed by changes in neurodegenerative biomarkers (CSF tau, F-18 fluorodeoxyglucose-positron emission tomography, magnetic resonance imaging) with the onset of clinical symptoms. The timing of these changes varies in individual patients due to genetic and environmental factors that increase or decrease an individual's resilience in response to progressive accumulations of AD pathologies. Further studies in ADNI will refine this model and render the biomarkers studied in ADNI more applicable to routine diagnosis and to clinical trials of disease modifying therapies. Copyright 2010 The Alzheimer
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                Author and article information

                Journal
                Proteomics Clin Appl
                Proteomics Clin Appl
                10.1002/(ISSN)1862-8354
                PRCA
                Proteomics. Clinical Applications
                John Wiley and Sons Inc. (Hoboken )
                1862-8346
                1862-8354
                25 March 2015
                April 2015
                : 9
                : 3-4 , Quantitation and Other Critical Issues in Clinical Proteomic Studies ( doiID: 10.1002/prca.v9.3-4 )
                : 406-422
                Affiliations
                [ 1 ] School of Life Sciences The University of Nottingham Nottingham UK
                Author notes
                [*] [* ] Correspondence: Dr. Lucy Fairclough, School of Life Sciences, The University of Nottingham, A Floor West Block, Queen's Medical Centre, Nottingham NG7 2UH, UK

                E‐mail: lucy.fairclough@ 123456nottingham.ac.uk

                Article
                PRCA1629
                10.1002/prca.201400130
                6680274
                25644123
                4b682d1b-52c3-4e15-abcf-e839cba6be89
                © 2015 The Authors. PROTEOMICS ‐ Clinical Applications Published by WILEY‐VCH Verlag GmbH & Co.

                This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc-nd/4.0/ License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made.

                History
                : 03 September 2014
                : 08 December 2014
                : 19 January 2015
                Page count
                Pages: 17
                Categories
                Review
                Reviews
                Custom metadata
                2.0
                prca1629
                April 2015
                Converter:WILEY_ML3GV2_TO_NLMPMC version:5.6.7 mode:remove_FC converted:05.08.2019

                Molecular biology
                elisa,in vitro diagnostics,multiplex immunoassay
                Molecular biology
                elisa, in vitro diagnostics, multiplex immunoassay

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