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Applying Proteome Partitioning to Biomarker Discovery in Plasma or Serum: Immunoaffinity Enrichment Strategies

Phenomenex Inc
10 May 2007

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Separation of complex mixtures that have an enormous range of protein concentration, such as plasma or serum, is a significant challenge for proteomic analysis. Specific removal of highly abundant proteins (HAP) during sample preparation can improve the ability to detect low abundance proteins (LAP) and enhance biomarker discovery.

Immunoaffinity partitioning of HAP has proven to be one of the most effective approaches for this purpose. Compared to IgG, avian IgY (Immunoglobulin Yolk) antibodies have unique and advantageous features that improve specific protein removal from primate, rodent and other mammalian samples.

This talk will describe the efficiency and effectiveness of the ProteomeLab™ IgY Proteome Partitioning Systems to separate 12 highly abundant proteins from plasma using immunoaffinity columns in spin column and liquid chromatography (LC) formats, and will present several new applications.

The results demonstrated high specificity of protein separation, with removal of 95-99.5% of the abundant human proteins, and the ability to dig substantially deeper into the proteome.

Phenomenex distributes Beckman Coulter’s innovative IgY proteome enrichment chemistry to advance the discovery of protein biomarkers

Phenomenex Inc., a leading worldwide supplier of separation science consumables, announced today the completion of a distribution agreement for Beckman Coulter's market-leading ProteomeLab™ IgY enrichment products for proteomics and biomarker discovery. The agreement grants Phenomenex exclusive distribution of the ProteomeLab IgY product line in North America and countries in the European Union. "Protein partitioning is a critical part of any biomarker discovery and we view the ProteomeLab IgY products as the best available," commented Michael McGinley, Phenomenex Biochromatography Product Manager. "ProteomeLab IgY fits well with our portfolio of separation products for proteins and peptides providing complete solutions for proteomics workflows."

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UHPLC and HPLCHigh performance liquid chromatography (HPLC) and ultra high performance liquid chromatography (UHPLC), also known as UPLC, are analytical techniques used to separate, identify and quantitate components of complex mixtures including biological samples such as proteins and lipids as well as chemical mixtures of pesticides, drugs and oils. Both techniques are liquid chromatographic methods but differ by operating pressures (HPLC < 6000 psi < UHPLC ). Components of HPLC and UHPLC systems include columns, detectors, pumps, autosamplers and column heaters. Explore a range of UHPLC and HPLC columns for your specific sample needs including reverse phase, normal phase, ion exchange, HILIC, ion exclusion and size exclusion columns. For more specialized HPLC, explore FPLC, countercurrent LC and simulated moving bed systems. Find the best UHPLC and HPLC equipment in our peer reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.AntibodiesAntibodies are used in techniques such as confocal and fluorescence microscopy, flow cytometry, ELISA, ELISPOT, immunohistochemistry, western blotting and immunopreciptation. Select specific antigen reactivity, high specific affinity, low non-specific binding, monoclonal or polyclonal, primary or secondary antibodies and associated conjugates such as an enzyme or dye for visualization.ProteomicsProteomics is the systemic bioinformatics study of proteins and amino acids, including their structure, size, function and identification. Tools used in proteomics include chromatography, blotting and gels, protein arrays, mass spectrometry and ELISA and associated analysis software. Analyzers and proteomic systems should be sensitive, high resolution, fast and may be automated for high-throughput.BiomarkersBiomarkers are biological markers which can be measured and evaluated to indicate a biological state. The use of biomarkers in research and diagnosis can indicate a normal or disease state or drug response of cells / tissues. Biomarkers include genetic markers, cell surface markers such as antigens, antibodies or receptors and secreted molecules such as cytokines. An assay system is required for identification of biomarkers. :

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Applying Proteome Partitioning to Biomarker Discovery in Plasma or Serum: Immunoaffinity Enrichment Strategies