EpiC: An Open Resource for Exploring Epitopes To Aid Antibody-Based Experiments
ABSTRACT Antibodies are a primary research tool for a diverse range of experiments in biology, from development to pathology. Their utility is derived from their ability to specifically identify proteins at a high level of sensitivity. This diversity of experimental requirements stretches the capabilities of these key research reagents. However, antibodies seem well placed to answer the challenges of the forthcoming proteome-scale biology. Their use in such a wide variety of experimental requirements impacts on the choice of epitope used to raise the antibody. Understanding the constraints imposed by the experimental configuration is crucial to developing well-characterized affinity reagents. Their application to a wide range of biological fields and relatively low-cost of manufacture has ensured that the demand for a resource of well-characterized antibodies will remain high and that they will be an important biological resource for the foreseeable future. This demand will only increase as the number of therapeutic targets continues to grow. Current tools to aid in the production of affinity reagents are disparate and not freely available. We present a freely available Web resource ( http://epic.embl.de ) for the proteomics community; the Epitope Choice Resource (EpiC) for the selection of epitopes and characterization of the target protein. It provides the community with a single Web-based portal for the exploration of epitopes on a target protein and connects over the Internet to a wide range of bioinformatic tools ensuring that data being presented are up to date.
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ABSTRACT: Bacterial infections represent a serious burden both for animal production and human health (zoonosis). Faster and more reliable diagnosis are mandatory in order to avoid economic losses and antibiotics misuse. The development of new potential diagnostic strategies for the immunodetection of pathogens is closely linked to the discovery of small polypeptides with immunogenic or immunoreactive activity. The candidate peptides used for this purpose must have several properties principally represented by their specificity and their location in the bacterial cell. Both proteomics, peptidomics and bioinformatics represent powerful complementary tools to discover specific immunoreactive peptides useful for diagnosis or vaccine. Peptidomics of Mycobacterium avium subsp. paratuberculosis (MAP) represents a good example of the potential of this discovery-phase. This review reports a comprehensive update of the current scientific knowledge about proteins and peptides of MAP with already documented humoral response. These findings, together with bioinformatics tools available, could be extremely useful to design a better strategy for subclinical bovine paratuberculosis diagnosis. The knowledge provided also represents a reliable example on the workflow to be followed in the direction of the diagnosis of other diseases through a peptidomic approach.04/2015; 2(1). DOI:10.1515/ped-2015-0001
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ABSTRACT: Alongside mass spectrometry, antibodies and other specific protein binding molecules have a special place in proteomics as affinity reagents in a toolbox of applications for determining protein location, quantitative distribution and function (affinity proteomics). The realisation that the range of research antibodies available, while apparently vast is nevertheless still very incomplete and frequently of uncertain quality, has stimulated projects with an objective of raising comprehensive, proteome-wide sets of protein binders. With progress in automation and throughput, a remarkable number of recent publications refer to the practical possibility of selecting binders to every protein encoded in the genome. Here we review the requirements of a pipeline of production of protein binders for the human proteome, including target prioritisation, antigen design, 'next generation' methods, databases and the approaches taken by ongoing projects in Europe and the USA. While the task of generating affinity reagents for all human proteins is complex and demanding, the benefits of well characterised and quality controlled pan-proteome binder resources for biomedical research, industry and life sciences in general would be enormous and justify the effort. Given the technical, personnel and financial resources needed to fulfil this aim, expansion of current efforts may best be addressed through largescale international collaboration.PROTEOMICS - CLINICAL APPLICATIONS 12/2013; 7(11-12). DOI:10.1002/prca.201300060 · 2.68 Impact Factor
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ABSTRACT: In affinity proteomics, specific protein-binding molecules (a.k.a. binders), principally antibodies, are applied as reagents in proteome analysis. In recent years, advances in binder technologies have created the potential for an unprecedented view on protein expression and distribution patterns in plasma, cells and tissues and increasingly on protein function. Particular strengths of affinity proteomics methods include detecting proteins in their natural environments of cell or tissue, high sensitivity and selectivity for detection of low abundance proteins and exploiting binding actions such as functional interference in living cells. To maximise the use and impact of affinity reagents, it will be essential to create comprehensive, standardised binder collections. With this in mind, the EU FP7 programme AFFINOMICS (http://www.affinomics.org), together with the preceding EU programmes ProteomeBinders and AffinityProteome, aims to extend affinity proteomics research by generating a large-scale resource of validated protein-binding molecules for characterisation of the human proteome. Activity is directed at producing binders to about 1000 protein targets, primarily in signal transduction and cancer, by establishing a high throughput, coordinated production pipeline. An important aspect of AFFINOMICS is the development of highly efficient recombinant selection methods, based on phage, cell and ribosome display, capable of producing high quality binders at greater throughput and lower cost than hitherto. The programme also involves development of innovative and sensitive technologies for specific detection of target proteins and their interactions, and deployment of binders in proteomics studies of clinical relevance. The need for such binder generation programmes is now recognised internationally, with parallel initiatives in the USA for cancer (NCI) and transcription factors (NIH) and within the Human Proteome Organisation (HUPO). The papers in this volume of New Biotechnology are all contributed by participants at the 5th ESF Workshop on Affinity Proteomics organised by the AFFINOMICS consortium and held in Alpbach, Austria, in March 2011.New Biotechnology 06/2012; 29(5):511-4. DOI:10.1016/j.nbt.2012.05.003 · 2.11 Impact Factor