2004-2008 Plan Meeting

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

HighGlycanPresentation 2013Outline of MP Bio PresentationIntroduction Scientific Portion: Surface chemistry of enzymes important for glycan sample preparationKits for glycoproteins extractionsNew FastPrep-5G instrumentOther possible forms of collaborations

HighGlycanPresentation 2013MP Bio New contact peopleDr. Daniel Weissbart; Dr. Vronique Karsten Dr. Kumar Bala, Dr. Jhony Orbulescu, Jeffrey Whyte

Financial/Legal: Mr. Barthel / Mr. Stankovitch.Lear: Mr. Fracois Barthel

Consultants: Prof. Miodrag Micic, Cerritos College & UCI Prof. Roger M. Leblanc, University of Miami. HighGlycanPresentation 2013Surface chemistry of enzymes for glycan sample prepFor 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 sensorsEnzyme currently studied: Beta Galactosidase, future work on l-fucosidase and fucose dehydrogenase and pngase HighGlycanPresentation 2013Langmuir monolayer studies of beta galactosidasesWhat is the Langmuir monolayer techniquePressure area isotherm slidePressure surface potential isothermSpectroscopic properties at monolayerFuture 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.

HighGlycanPresentation 2013What is the Langmuir Monolayer ? 5

A one-molecule thick layer of an insoluble or amphiphilic material spread onto water/aqueous subphaseWorks for most proteins and enzymes.Can be used in a form of the mixed colayersCan be transferred to a solid substrate surface as a film,a.k.a Langmuir Blodgett filmIn vitro model of biological membraneControllable experimental variablesEfficient heterogenous catalysts and sensorsHighGlycanPresentation 2013Langmuir Monolayer Technique6AirWaterAirWaterAirWater

HighGlycanPresentation 2013Langmuir-Blodgett technique - transfeering highly packed enzyme monolayer to the solid substrate7

HighGlycanPresentation 2013Current Lanmguir Monolayer results on beta galactosidaseAs a model system we have performed a study of the beta galactosidase from e coli (MP 0215003901)Obtained stable and reproducible surface-pressure-area isothermsStudied spectroscopic properties in the lanmguir monolyerCurrent work: Formation of stable isotherms with Mg2+ subphaseMonolayer transfer to hydrophilic and hydrophobic substrates.

8HighGlycanPresentation 2013Beta Galactosidase from e coli (MP0214003901)-Galactosidase cleaves terminal galactose residues, which are -1-->14 linked to a monosaccharide, glycopeptide, or oligosaccharide. The enzyme requires Mg2+ for optimal activity. -galactosidase cleaves lactose into its monosaccharide components, glucose and galactose. It also catalyses the transglycosylation of glucose into allolactose, the inducer of -galactosidase.9HighGlycanPresentation 2013Spectroscopic properties of beta galactosidase, bulk10

HighGlycanPresentation 2013Surface pressure area isotherms of Beta Galactosidease with estimated average mean molecular surface areas at the 2D solid state film.

HighGlycanPresentation 2013Both spectroscopic and surface pressure area data indicates stable 2D monolayer solid film 12

HighGlycanPresentation 2013Finalizing 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 filmsSecond manuscript on LB film transfer and properties. 13Beta Galactosidase current work:HighGlycanPresentation 2013Fucose Dehydrogenase, Fucosidase and PNGase surface chemistry workCurrently working on souricing/ recombinant expression of Fucose Dehydrogenase, L-Fucosidase and PNGaseLanngmuir, Langmuir Blodgett studies individual enzymesEnzyme mixtures Practical applications:Enzyme mixture for glycan release in the form of Heterogenous catalysis / immobilized sample prep labwareSecondary outcome: MP Bio will be able to provide low cost supply of highpurity enzymes and enzyme mixtures once the expression is established HighGlycanPresentation 2013Glycoproteins sample prep kit worksKits for glycoprotein extractionsLectin ConA based columns for purification of N-linked glycoprotein fractions using the FastPrep Lysis Simplified workflow, bind-wash-elute WGA and ConA columnProcedure : FastPrep LysisBind; Wash EluteFirst samples available for testing Nov-Dec 2013.

HighGlycanPresentation 2013Key Features 16FastPrep-24-5G : Next Generaton FastPrep 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 Predefined Settings, MP RecommendedWill include settings for FastGlycoProtein kitsManual SettingsStore up to 12 User-defined Settings (from 5)Export DataRe-order and tech serve info for 19 global regionsBluetooth CompatableUSB Port, App UpdatesVoice Interface that will give run statisticsStrobe Light to aid visual lysis inspection

HighGlycanPresentation 2013Other collaboration with Consoritum ParticipantsMP bio is an affordable high quallity reagents supply provider90% of Sigma Aldrich offerings High quallity reagents, kits and systems:BiochemicalsRadiochemicalsImmunologicalsMolecular Biology RIA assaysFine chemical and biologicals sourcing and custom manufacturing group for custom products. HighGlycanPresentation 2013Who 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.HighGlycanPresentation 201318HistoryFormerly 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 17 000 employees.2003, ICN is sold.2004, Milan Panic founded MP Biomedicals through the acquisition of thirty companies, including Qbiogen. HighGlycanPresentation 20131920

Santa Ana, CAAmerican Headquarter

Solon, OHFacilities and logistic

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Sydney, AutraliaFacilities and logisticsMP Biomedicals in the world

Eschwege GermanyFacilities and LogisticsHighGlycanPresentation 20132021Our ApplicationsLife ScienceDiagnosticFine ChemicalsHighGlycanPresentation 20132122LifeScienceFrom 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 preparationCell BiologyMolecular BiologyProteomicsDrug DiscoverySolid Phase Peptide Synthesis

HighGlycanPresentation 20132223ChemicalsMP 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 ReagentsBuilding BlocksProtecting &Deprotecting ReagentsReduction &Oxidation ReagentsPrecious Metal compounds and catalystsRare Earths Amino AcidsSilica & Alumina / ChromatographyAlbuminsAbsorbentsAnd moreHighGlycanPresentation 20132324DiagnosticWhether 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 hoursAuto-immunes KitsBiogenic AminesCardiac MarkersEndocrinologyFood QualityInfectious DiseasesIsocalibratorsNewborn ScreeningCancer MarkersHighGlycanPresentation 201324

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HighGlycanPresentation 201325THANK YOU ! Q & A ?

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