www.proteocell.comBIOPROCESS AT PROTEOCELLMolecularEngineeringProcessEngineeringFacilityEngineering
www.proteocell.comMolecularEngineeringExpression plasmid designand generation(Mammalian, Yeast, and E.Coli cell systems)Sequence Analysis,construct design, andmolecular optimizationUpstream Process(Cell Culture)Downstream Process(Purification)Transient transfectionClonal selectionGeneration of optimal stable cell lineCreation of Master and Working CellbanksSmall & Medium scale cell culture(Mammalian, Yeast, & E. Coli)Molecular engineeringexpertise & consultingBioprocess Scale-up expertise & consultingPurification expertise &consultingGLP & GMP Material ProductionBioprocess Analytics suite—process and auxiliary bio-analytics servicesSmall & Medium scalePurificationViral ClearanceUltrafiltration/Dialfiltration/ConcentrationPurified end-productsNon-purified end products (clarified harvest)Small-scale bioprocess development & optimization (CellCulture & Purification)Cell Culture & fermentationexpertise & consultingPROTEOCELL BIOTECHNOLOGIES’ SPECTRUM OF SERVICES
www.proteocell.com3 KEYS TO EFFECTIVE BIOPROCESS FOR BIOPHARMA MANUFACTURING1) Superior cell line and cell line technology used;2) Efficient and effective bioprocess development;3) A-Z adequate regulatory compliance.
www.proteocell.comBIOPROCESS INTEGRATION STRATEGY OF PROTEOCELL SSI1) Cell Line Engineering:•Using the latest in Ab molecular engineering methods to design a DNA expression vectorsystem that is stable and high producing, and scalable to a production system (bioreactorscale)2) Process Development:• Preliminary bioprocess work that ProteoCell has successfully incorporated into its cell lineengineering strategy by overlapping clonal selection concepts with Process Developmentconcepts;• It includes the preliminary expression of the designed Ab clones in a quasi-production/fullproduction setting to test for product homogeneity and stability—before final clonal selectionand cell banking;• This includes production in a bioreactor, simulating production batch conditions, includingchemically defined media and other regulatory compliant bioprocess conditions.
www.proteocell.comMOLECULAR ENGINEERING HIGHLIGHTS OF PROTEOCELL SSI SYSTEM1) Alternate “tagging-targeting vector” mechanism for rapid generationof stable genomic integration of target gene sequence;2) Strong CMV promoter drives LC or HC expression;3) Weaker promoter or IRES drives selectable marker to ensure thatMAb expression is dominant;4) Single or double fluorescence tagging of protein sequence dependingon experimental design strategy.
www.proteocell.comPROTEOCELL SSI = SPEED; CLONAL SELECTION IN RECORD TIME!!Targeting vectorCulture of appropriatetagged cell lineWeeks 1,2 Weeks 2,3 Weeks 4,5Clonal selection and expressionTransfection
www.proteocell.comPRELIMINARY PROCESS DEV. OF A CELL LINE USING PROTEOCELL SSI PLATFORM•Initial Process Development in Flask (Batch and Fed Batch mode) and further optimizationin Bioreactor culture.•Multiple tracks are then undertaken for optimizing clonal conditions in both flasks andbioreactor culture (before MCB is produced to confirm scale-up capability of selectiveclone)Media/Culture ParametersCellGrowthCellViabilityCultivationTime01234567Productivity(FoldIncrease)Preliminary Process Dev. ProjectionFlask Process Dev.Months 1 2 3 4Flask /Bioreactor Process Dev.
www.proteocell.comINCREASED ANTIBODY YIELD IN CHO CELLS WITH PROTEOCELL SSI PLATFORM0123456IgGTiter(g/L)Cell Line Engineering 30-60 daysPreliminary Process Dev. 30-75 daysAdvanced Process Dev. 45-75 days
www.proteocell.comPROJECTING INTO PROCESS DEVELOPMENT AND BEYOND....Completion of Cell line engineering and Preliminary Process DevelopmentGMP Cell Banking and Process DevelopmentProcess Scale-upPilot scale cGMP process development at ProteoCell (Engineering runs)Tech transfer to commercial manufacturing facility
www.proteocell.comTHE PROTEOCELL SSI MFG. PROCESS STAGES OF DEVELOPMENT OUTLINEStage 2: Process Scale-up to50-100L scaleStage 1: GMP Cell Line andProcess DevelopmentStage 3: GMP Pilot scaleprocess productionStage 4: Technology Transferto Commercial Mfg. site3 90 6 12 15 18 21 24(months)