Presentation on theme: "“PAT” Applications for Biochemical Processes"— Presentation transcript:
1 “PAT” Applications for Biochemical Processes Shih-Hsie PanInterphex, March 19, 2009
2 PAT Framework Multivariate data acquisition and analysis tools Process chemometricsIntelligent use of process dataModern process analyzersProcess analytical chemistry toolsIn-process monitoring techniquesProcess AnalysisProcess MonitoringProcess ControlProcess DesignProcess and endpoint monitoring and control toolsProcess supervisory controlHigh level multivariate control strategiesDesign for QualityContinuous improvement and knowledge management toolsFMECADOE
3 PAT: Process Information Enabling QbD LaboratoryProductionAreaDivertedSampleInsertedProbeNo ProductContactOff-LineAt-LineOn-LineIn-LineNonInvasiveReal-timereleaseTransition AnalysisPredictive ModelingNIR Probe
4 Benefits of PAT for Biologics Increase knowledge of product and processIdentify critical steps and parameters (CCP’s and CPP’s) that impact qualityLower the cost of process improvement to increase yield, quality & robustnessMinimize process validation cost – direct, real-time process controlFacilitate reduction of batch-to-batch variability for better quality and predictabilityAllow near real time critical parameter conformance monitoring and comparisons – continuous quality assurance and validationAssist validation efforts for characterization and documentation of process changesReduce testing requirements at end of processAssess deviation impact in real timeAvoid costs of processing unreleasable batchesData justification of batch releaseProvide an ability to quickly identify shifts, trends, or outliers in the data, so that investigations can be conducted and decisions made on lot release quickly to reduce manufacturing risk.
5 Significant Reduction in RSD Improved Consistency in Automated (At-Line) Cell Count and Viability DeterminationBy Image AnalysisSignificant Reduction in RSD Improved Consistency inMfg Operations based on Cell Count or %ViabilityCourtesy of Polina Rapoport
6 Chromatographic Transition Analysis Real time method developed for monitoring column packing quality.Calculates plate number directly from transition curve.No off-line pulse injection tests required; uses process data.Predictive of column performance.
7 Affinity Elution Chromatogram Loss of Column IntegrityAffinity Elution ChromatogramChromatogram improved after lowering flow adapter
8 Transition Analysis Identifies Changes Column RepackedLowered Flow AdapterHETP data clearly identifies changes in column integrity.Values increase with time after column packing.Original HETP value is restored after lowering the top flow adapter.Increased measurement variability is observed when column integrity decreases.
9 Packed Cell VolumePCV is an accurate measurement of biomass, but it also lends itself to many inconsistencies…1) Manual operation that is variable from operator to operator.2) Measurement is performed visually which can also be very subjective.Drivers to evaluate alternative methods of determining biomass to ensure a more robust and informative estimate of inoculum transfer time.
10 Oxygen Transfer Rate (OTR) DefinitionkLa = mass transfer coefficient ,based on empirical data from each bioreactor familyC* = dissolved oxygen level at oxygen saturation pointCL = Dissolved Oxygen Concentration (should be a constant)Pros-OTR directly measures cell growthOTR is a non-invasive method, per guidance definition
11 Using Technology…. To manage process performance Case Study ResultsR2 Value vs. Current(Off-Line) MethodOn-LineMethodNon-Invasive MethodN-3 Stage0.920.91N-2 Stage0.970.95N-1 Stage0.84Using Technology…. To manage process performance
12 Data courtesy of Kirin Jamison Prediction of protein titers with PLS modelbased on 1695 variablesData courtesy of Kirin Jamison
13 Acknowledgement My colleagues at Genentech: Eric Fallon Robert Kiss Harry Lam
15 Amino acid analysis by on-line HPLC Additional At-Line Analyses Have Increased Measurable ParametersBlood gas analyzersEnable measurement of glucose, lactate, pCO2, pH, pO2, ammonium, sodium, potassium and other metabolitesAmino acid analysis by on-line HPLCAmino acids along with glucose can be measured every hour with automated HPLCCan enable more comprehensive view of how metabolism shifts over the course of a cultureCan also be used for medium development & optimizationAutomated image analysis for cell count, viability, cell size (example)