Presentation on theme: "0 Current Progress on Advanced Systems for Glycomics Sample Preparation Split, 10-2-2013. Illkirch, June 2011."— Presentation transcript:
0 Current Progress on Advanced Systems for Glycomics Sample Preparation Split, Illkirch, June 2011
HighGlycan Presentation 2013 Outline of MP Bio Presentation uIntroduction uScientific Portion: Surface chemistry of enzymes important for glycan sample preparation uKits for glycoproteins extractions uNew FastPrep-5G instrument uOther possible forms of collaborations
HighGlycan Presentation 2013 MP Bio New contact people uDr. Daniel Weissbart; Dr. Véronique Karsten uDr. Kumar Bala, Dr. Jhony Orbulescu, Jeffrey Whyte uFinancial/Legal: Mr. Barthel / Mr. Stankovitch. uLear: Mr. Fracois Barthel Consultants: uProf. Miodrag Micic, Cerritos College & UCI uProf. Roger M. Leblanc, University of Miami.
HighGlycan Presentation 2013 Surface chemistry of enzymes for glycan sample prep uFor the first time studied behavior of enzymes important for the glycan preparation in the Langmuir monolayer. uFuture work: Langmuir blodgett film enzyme immobilization for the purpose of creating an efficient heterogenous support for columns and possibly sensors uEnzyme currently studied: Beta Galactosidase, future work on l-fucosidase and fucose dehydrogenase and pngase
HighGlycan Presentation 2013 Langmuir monolayer studies of beta galactosidases uWhat is the Langmuir monolayer technique uPressure area isotherm slide uPressure surface potential isotherm uSpectroscopic properties at monolayer uFuture work on the enzyme immobilization in the form of the Langmuir Blodgett film uPractical application: heterogenous bio catalyst for glycans cleavage/sample preparation based on immobilized Lagmuir Blodgett films on substrate.
HighGlycan Presentation 2013 What is the Langmuir Monolayer ? 5 1.A one-molecule thick layer of an insoluble or amphiphilic material spread onto water/aqueous subphase 2.Works for most proteins and enzymes. 3.Can be used in a form of the mixed colayers 4.Can be transferred to a solid substrate surface as a film,a.k.a Langmuir Blodgett film 5.In vitro model of biological membrane 6.Controllable experimental variables 7.Efficient heterogenous catalysts and sensors
HighGlycan Presentation 2013 Langmuir Monolayer Technique 6 Air Water Air Water Air Water
HighGlycan Presentation 2013 Langmuir-Blodgett technique - transfeering highly packed enzyme monolayer to the solid substrate 7
HighGlycan Presentation 2013 Current Lanmguir Monolayer results on beta galactosidase uAs a model system we have performed a study of the beta galactosidase from e coli (MP ) uObtained stable and reproducible surface-pressure- area isotherms uStudied spectroscopic properties in the lanmguir monolyer uCurrent work: Formation of stable isotherms with Mg2+ subphase uMonolayer transfer to hydrophilic and hydrophobic substrates. 8
HighGlycan Presentation 2013 Beta Galactosidase from e coli (MP ) uβ-Galactosidase cleaves terminal galactose residues, which are β -1-->14 linked to a monosaccharide, glycopeptide, or oligosaccharide. The enzyme requires Mg2+ for optimal activity. uβ-galactosidase cleaves lactose into its monosaccharide components, glucose and galactose. It also catalyses the transglycosylation of glucose into allolactose, the inducer of β- galactosidase. 9
HighGlycan Presentation 2013 Surface pressure area isotherms of Beta Galactosidease with estimated average mean molecular surface areas at the 2D solid state film.
HighGlycan Presentation 2013 Both spectroscopic and surface pressure area data indicates stable 2D monolayer solid film 12
HighGlycan Presentation 2013 uFinalizing first manuscript/communications on surface properties of monolayer of beta galactosidase. Intended submission: Colloids & Surfaces B:Biointerfaces or Langmuir. uMake reproducible surface pressure area istoherm with Mg2+ active subphase. uTransfeer Langmuir Blodgett film to mica and glass substrates uTest stability and activity of the solid state LB films uSecond manuscript on LB film transfer and properties. 13 Beta Galactosidase current work:
HighGlycan Presentation 2013 Fucose Dehydrogenase, Fucosidase and PNGase surface chemistry work Currently working on souricing/ recombinant expression of Fucose Dehydrogenase, L-Fucosidase and PNGase Lanngmuir, Langmuir Blodgett studies individual enzymes Enzyme mixtures Practical applications: Enzyme mixture for glycan release in the form of Heterogenous catalysis / 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
HighGlycan Presentation 2013 Glycoproteins sample prep kit works uKits for glycoprotein extractions uLectin ConA based columns for purification of N-linked glycoprotein fractions using the uFastPrep Lysis uSimplified workflow, bind-wash-elute uWGA and ConA column uProcedure : uFastPrep Lysis uBind; uWash uElute uFirst samples available for testing Nov-Dec 2013.
HighGlycan Presentation 2013 Key Features 16 FastPrep-24-5G : Next Generaton FastPrep uSpeed Range: M/S (from 6.5) uTime: Seconds (from 60) uCycles: 1-9 (New feature) uPause between cycles: seconds (New feature) uNoise: <60 dB uTouchscreen User Interface (all new) u>60 Predefined Settings, MP Recommended uWill include settings for FastGlycoProtein kits uManual Settings uStore up to 12 User-defined Settings (from 5) uExport Data uRe-order and tech serve info for 19 global regions uBluetooth® Compatable uUSB Port, App Updates uVoice Interface that will give run statistics uStrobe Light to aid visual lysis inspection
HighGlycan Presentation 2013 Other collaboration with Consoritum Participants uMP bio is an affordable high quallity reagents supply provider u90% of Sigma Aldrich offerings uHigh quallity reagents, kits and systems: uBiochemicals uRadiochemicals uImmunologicals uMolecular Biology uRIA assays uFine chemical and biologicals sourcing and custom manufacturing group for custom products.
HighGlycan Presentation 2013 Who we are MP Biomedicals is 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.
HighGlycan Presentation 2013 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 is founded 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 largest shareholder of Roche. 2002, ICN listed as 49th largest pharmaceutical company in the world with employees. 2003, ICN is sold. 2004, Milan Panic founded MP Biomedicals through the acquisition of thirty companies, including Qbiogen.
HighGlycan Presentation Santa Ana, CA American Headquarter Solon, OH Facilities and logistic Strasbourg, France European Headquarter Singapore Asia Headquarter Sydney, Autralia Facilities and logistics MP Biomedicals in the world Eschwege Germany Facilities and Logistics
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HighGlycan Presentation 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. Sample preparation Cell Biology Molecular Biology Proteomics Drug Discovery Solid Phase Peptide Synthesis
HighGlycan Presentation 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. Specials Synthetic Reagents Building Blocks Protecting &Deprotecting Reagents Reduction &Oxidation Reagents Precious Metal compounds and catalysts Rare Earths Amino Acids Silica & Alumina / Chromatography Albumins Absorbents And more…
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HighGlycan Presentation 2013 THANK YOU ! Q & A ? 26