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Current Progress on Advanced Systems for Glycomics Sample Preparation Split, 10-2-2013.

Current Progress on Advanced Systems for Glycomics Sample Preparation Split, 10-2-2013. Illkirch, June 2011. Outline of MP Bio Presentation. Introduction Scientific Portion: Surface chemistry of enzymes important for glycan sample preparation Kits for glycoproteins extractions

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Current Progress on Advanced Systems for Glycomics Sample Preparation Split, 10-2-2013.

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  1. Current Progress on Advanced Systems for Glycomics Sample PreparationSplit, 10-2-2013. Illkirch, June 2011

  2. Outline of MP Bio Presentation Introduction Scientific Portion: Surface chemistry of enzymes important for glycan sample preparation Kits for glycoproteins extractions New FastPrep-5G instrument Other possible forms of collaborations

  3. MP Bio New contact people Dr. Daniel Weissbart; Dr. Véronique Karsten Dr. Kumar Bala, Dr. JhonyOrbulescu, Jeffrey Whyte Financial/Legal: Mr. Barthel / Mr. Stankovitch. Lear: Mr. FracoisBarthel Consultants: Prof. Miodrag Micic, Cerritos College & UCI Prof. Roger M. Leblanc, University of Miami.

  4. Surface chemistry of enzymes for glycan sample prep For the first time studied behavior of enzymes important for the glycan preparation in the Langmuir monolayer. Future work: Langmuir blodgett film enzyme immobilization for the purpose of creating an efficient heterogenous support for columns and possibly sensors Enzyme currently studied: Beta Galactosidase, future work on l-fucosidase and fucose dehydrogenase and pngase

  5. Langmuir monolayer studies of beta galactosidases What is the Langmuir monolayer technique Pressure area isotherm slide Pressure surface potential isotherm Spectroscopic properties at monolayer Future work on the enzyme immobilization in the form of the Langmuir Blodgett film Practical application: heterogenous bio catalyst for glycans cleavage/sample preparation based on immobilized Lagmuir Blodgett films on substrate.

  6. What is the Langmuir Monolayer ? A one-molecule thick layer of an insoluble or amphiphilic material spread onto water/aqueous subphase Works for most proteins and enzymes. Can be used in a form of the mixed colayers Can be transferred to a solid substrate surface as a film,a.k.a Langmuir Blodgett film In vitro model of biological membrane Controllable experimental variables Efficient heterogenous catalysts and sensors

  7. Langmuir Monolayer Technique Air Water Air Water Air Water

  8. Langmuir-Blodgett technique - transfeering highly packed enzyme monolayer to the solid substrate

  9. Current Lanmguir Monolayer results on beta galactosidase As a model system we have performed a study of the beta galactosidase from e coli (MP 0215003901) Obtained stable and reproducible surface-pressure-area isotherms Studied spectroscopic properties in the lanmguirmonolyer Current work: Formation of stable isotherms with Mg2+ subphase Monolayer transfer to hydrophilic and hydrophobic substrates.

  10. Beta Galactosidase from e coli (MP0214003901) β-Galactosidase cleaves terminal galactose residues, which are β -1-->14 linked to a monosaccharide, glycopeptide, or oligosaccharide. The enzyme requires Mg2+ for optimal activity. β-galactosidase cleaves lactose into its monosaccharide components, glucose and galactose. It also catalyses the transglycosylation of glucose into allolactose, the inducer of β-galactosidase.

  11. Spectroscopic properties of beta galactosidase, bulk

  12. Surface pressure area isotherms of Beta Galactosidease with estimated average mean molecular surface areas at the 2D solid state film.

  13. Both spectroscopic and surface pressure area data indicates stable 2D monolayer solid film

  14. Beta Galactosidase current work: Finalizing first manuscript/communications on surface properties of monolayer of beta galactosidase. Intended submission: Colloids & Surfaces B:Biointerfaces or Langmuir. Make reproducible surface pressure area istoherm with Mg2+ active subphase. Transfeer Langmuir Blodgett film to mica and glass substrates Test stability and activity of the solid state LB films Second manuscript on LB film transfer and properties.

  15. FucoseDehydrogenase, Fucosidase and PNGase surface chemistry work • Currently working on souricing/ recombinant expression of FucoseDehydrogenase, L-Fucosidase and PNGase • Lanngmuir, Langmuir Blodgett studies • individual enzymes • Enzyme mixtures • Practical applications: • Enzyme mixture for glycan release in the form of • Heterogenouscatalysis / immobilized sample prep “labware” • Secondary outcome: MP Bio will be able to provide low cost supply of high • purity enzymes and enzyme mixtures once the expression is established

  16. Glycoproteins sample prep kit works • Kits for glycoprotein extractions • LectinConA based columns for purification of N-linked glycoprotein fractions using the • FastPrepLysis • Simplified workflow, bind-wash-elute • WGA and ConA column • Procedure : • FastPrepLysis • Bind; • Wash • Elute • First samples available for testing Nov-Dec 2013.

  17. FastPrep-24-5G : Next GeneratonFastPrep • Touchscreen User Interface (all new) • >60 Predefined Settings, MP Recommended • Will include settings for FastGlycoProtein kits • Manual Settings • Store up to 12 User-defined Settings (from 5) • Export Data • Re-order and tech serve info for 19 global regions • Bluetooth® Compatable • USB Port, App Updates • Voice Interface that will give run statistics • Strobe Light to aid visual lysis inspection • Speed Range: 4.0-10.0 M/S (from 6.5) • Time: 1-120 Seconds (from 60) • Cycles: 1-9 (New feature) • Pause between cycles: 1-300 seconds (New feature) • Noise: <60 dB Key Features 16

  18. Other collaboration with Consoritum Participants • MP bio is an affordable high quallity reagents supply provider • 90% of Sigma Aldrich offerings • High quallity reagents, kits and systems: • Biochemicals • Radiochemicals • Immunologicals • Molecular Biology • RIA assays • Fine chemical and biologicals sourcing and custom manufacturing group for custom products.

  19. Whowe are MP Biomedicalsis a global company committed in the development, manufacturing and marketing of life sciences and diagnostic products. The company is dedicated to provide scientists and researchers innovative and high performance tools combined with superior service to help them in their quest for discovery and thus make the dream of a better life, a reality.

  20. History Formerly ICN Pharmaceuticals, MP Biomedicals was formed in 2004 after the acquisition of various companies in various fields of application and complementary. 1959, ICN isfounded by Milan Panic. 1970, ICN discovers the first broad-spectrum antiviral drug in the world. Sales reach $ 1.5 billion and several million people were treated. 1986, ICN was the second largestshareholder of Roche. 2002, ICN listed as 49th largestpharmaceuticalcompany in the world with 17 000 employees. 2003, ICN issold. 2004, Milan Panic founded MP Biomedicals through the acquisition of thirty companies, including Qbiogen.

  21. MP Biomedicalsin the world Strasbourg, France EuropeanHeadquarter Eschwege Germany Facilities and Logistics Santa Ana, CA American Headquarter Singapore AsiaHeadquarter Solon, OH Facilities and logistic Sydney, Autralia Facilities and logistics

  22. Our Applications Life Science Fine Chemicals Diagnostic

  23. LifeScience From molecular biology, cell biology through apoptosis and immunology products MP BIOMEDICALS accompanies you at every stages of your research in life sciences. Many researchers have chosen our products because they guarantee returns and meet your requirements. We now supply both large universities, as well as major industries that quests reliable and innovative products. Samplepreparation CellBiology MolecularBiology Proteomics Drug Discovery Solid Phase Peptide Synthesis

  24. Chemicals MP Biomedicals provides more than 12,000 compounds of the K & K library. More than 1,000 of them are exclusive to MP Biomedicals and can be found in our product catalog. These compounds demonstrate our commitment to provide researchers with the largest selection of rare and fine chemicals, many of which are available nowhere else. Our wide range of compounds includes a variety of functional groups, both aromatic and nonaromatic. In this collection, you will find chiral compounds, inorganic rare earth compounds, amino acids, organometallic compounds and more. We also have a wide range of silica and alumina for chromatography. SpecialsSyntheticReagents Building Blocks Protecting &DeprotectingReagents Reduction &OxidationReagents PreciousMetal compounds and catalysts Rare Earths AminoAcids Silica & Alumina / Chromatography Albumins Absorbents And more…

  25. Diagnostic Whether you work with assays for animal testing or products geared toward human specimens, we are committed to provide you with kits and reagents of superior quality. Easy to use for determining in vitro concentration of various hormones in human and mouse samples. Each kit contains all necessary reagents, including controls, and produces accurate results within 24 hours Auto-immunes Kits BiogenicAmines CardiacMarkers Endocrinology Food Quality InfectiousDiseases Isocalibrators Newborn Screening Cancer Markers

  26. TradeShows Examples of Europeans Events

  27. Q & A ? THANK YOU !

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