Applying QbD to Biotech Process Validation
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The document outlines an experiment conducted by Biotech Logic Inc. focused on applying Quality by Design (QbD) principles to the validation of a biotechnology process utilizing ion exchange chromatography. It details the parameters evaluated, risk assessments, and experimental designs employed to ensure acceptable product yield and quality. The study concludes that the modified process design meets predefined specifications and has successfully confirmed the expected outcomes of the validation study.
Applying QbD to Biotech Process Validation
- 1. BIBioTech BI Applying Validation A case stu Stage 1 – TECH LOGIC BioTechLogic, Inc. BIO TECH LOGICBIO ® TECH IOLOGIC Logic, Inc. IO ® g QbD to Biotech Process n : udy in applying QbD to – Process Design IVT Validation Week 28-30 March 201128 30 March 2011 Kurtis Epp, John Kandl BioTechLogic, Inc.
- 2. Agenda • Process Backgrou • Risk Assessment • DOE • Parameter Evaluat • Conclusions TECH LOGICBIO TECH LOGICBIO ®BioTechLogic, Inc. und tion Slide 2 Company Confidential
- 3. Process Back The scope of this case stup data obtained from small- Chromatography runs per limits in experiments planlimits in experiments plan Design of Experiments (D TECH LOGICBIO TECH LOGICBIO ®BioTechLogic, Inc. kground udy is to provide and evaluatey p -scale Ion Exchange rformed within defined parameter ned and executed according toned and executed according to DOE). Slide 3 Company Confidential
- 4. Process Back TECH LOGICBIO TECH LOGICBIO ®BioTechLogic, Inc. kground Slide 4 Company Confidential
- 5. Risk Assessm Operating Parameter Normal Operating UV at Start of AEX Fraction Collection > 0.50 AU UV at End of AEX Fraction Collection < 20% of maximum p Elution Gradient 8% per CV Elution Gradient (20 – 100% B ove Equilibration volume ≥ 5 CV Load flow rate 100 – 120 L/h (Targe Wash flow rate 100 – 120 L/h (Targe Elution flow rate 100 – 120 L/h (Targe Pre-Equilibration flow rate 100 – 120 L/h (Targe F1 F3: 0 2 Fraction Volume F1 – F3: 0.2 F4+: 0.1 CV Wash volume ≥2 CV Fraction Mixing Speed 65 rpm Fraction Mixing Time 20 - 30 min Pool Mixing Time 10 – 15 min Pool Mixing Time Pool Mixing Speed 100 rpm Pre-Equilibration volume ≥ 4 CV WFI Rinse volume ≥ 3 CV WFI Rinse flow rate 100 – 120 L/h (Targe Equilibration flow rate 100 – 120 L/h (Targeq ( g In-Process Control Limits Column Load 15 – 25 g / L r Column Bed Height 30 ± 3 cm Column Backpressure during Equilibration, Load, Wash, and Elution < 3 bar Effl t H t d f E ilib ti ± 0.3 pH Units of Eq Effluent pH at end of Equilibration p q Buffer pH Effluent Cond. at end of Equilibration ± 1 mS/cm Units of E Buffer Conduc Effluent UV at the end of Equilibration Zero Fraction Pooling Criteria (RP-HPLC) ≥ 95% Main P TECH LOGICBIO TECH LOGICBIO ®BioTechLogic, Inc. ment g Range Potential Effect(s) of Failure SEV OCC DET RPN U Product loss, quality 9 4 2 72 peak height Product loss, quality 9 4 2 72 V Inconsistent quality 8 2 4 64 r 10 CV) Inconsistent quality 8 2 4 64 Product loss, quality 9 2 1 18 et: 110 L/h) Inconsistent quality 4 1 3 12 et: 110 L/h) Inconsistent quality 4 1 3 12 et: 110 L/h) Inconsistent quality 4 1 3 12 et: 110 L/h) Longer equilibration 4 1 3 12 CVCV V Product loss, quality 9 1 1 9 Inconsistent quality 5 1 1 5 Inconsistent quality 4 1 1 4 n Inconsistent quality 4 1 1 4 n Non-homogeneity, inconsistent sampling/yield 4 1 1 4 sampling/yield Non-homogeneity, inconsistent sampling/yield 4 1 1 4 Longer equilibration 3 1 1 3 Longer equilibration 3 1 1 3 et: 110 L/h) Longer equilibration 1 1 1 3 et: 110 L/h) Product loss, quality 3 1 1 3) , q y resin m quilibrationq Equilibration ctivity Peak Slide 5 Company Confidential
- 6. Risk Assessm The parameters selectedp the UV at start of fraction collection, gradient slope D d t i blDependent variables ana and purity by RP-HPLC. Testin Input parameter Lower UV at the beginning of fraction collection (AU) 0.40 UV at the end of pooling (% from peak max) 10 Gradient slope (% per CV) 4.0 Resin Load, (g/L resin) 15 TECH LOGICBIO TECH LOGICBIO ®BioTechLogic, Inc. ment d for evaluation in this study werey n collection, UV at end of fraction e, and resin load. l d t i i ldalyzed were protein recovery yield ng limitsg Output parameters Upper 0.60 Purity by RP-HPLC (≥ 95%) Step yield (≥ 75%) 30 12.0 25 Slide 6 Company Confidential
- 7. DOE Pattern Exp. No.: UV at the sta of fraction collection p collection (AU) −−−− 1 0.4 0000 2 0.5 0000 3 0 50000 3 0.5 −++− 4 0.4 0000 5 0.5 ++−− 6 0.6 7 0 6+−−+ 7 0.6 ++++ 8 0.6 +−+− 9 0.6 −−++ 10 0.4 0000 11 0.5 −+−+ 12 0.4 Note: It is important to randomize independence of your observation mistake by the operator. TECH LOGICBIO TECH LOGICBIO ®BioTechLogic, Inc. art n UV at the end of pooling Gradient slope Resin Load (g/L) (% peak max) (% per CV) (g/L) 10 4 15 20 8 20 20 8 2020 8 20 30 12 15 20 8 20 30 4 15 10 4 2510 4 25 30 12 25 10 12 15 10 12 25 20 8 20 30 4 25 e the run order to assure the ns and reduce the chances of a Slide 7 Company Confidential
- 8. Experimental • A summary of the var Ion Exchange Chrom Outputs step yield anOutputs, step yield an listed in the following • Runs that did not mee are shaded to indicate TECH LOGICBIO TECH LOGICBIO ®BioTechLogic, Inc. Results riable parameters for each of the atography runs as well as the nd eluate purity by RP HPLC arend eluate purity by RP-HPLC are table. et the defined acceptance criteriap e run failure Slide 8 Company Confidential
- 9. Experimental Run Resin Load GradientRun Number Resin Load (g/L) Gradient Slope (%) 1 15 4 2 20 8 3 20 8 4 15 12 5 20 85 20 8 6 15 4 7 25 4 8 25 128 25 12 9 15 12 10 25 12 11 20 811 20 8 12 25 4 TECH LOGICBIO TECH LOGICBIO ®BioTechLogic, Inc. Results UV @ Start % Peak Step Yield (%) RP-HPLC Purity (%)UV @ Start Collection Height End Collection (%) Purity (%) Acceptance Criteria ≥ 45% ≥ 95% 0.4 10 53 95 0.5 20 56 97 0.5 20 50 97 0.4 30 59 96 0 5 20 51 970.5 20 51 97 0.6 30 53 97 0.6 10 52 96 0 6 30 40 970.6 30 40 97 0.6 10 50 98 0.4 10 45 97 0.5 20 51 970.5 20 51 97 0.4 30 60 97 Slide 9 Company Confidential
- 10. Statistical dat • For evaluation of statisti on protein quality and q software package was u Squares model for EffecSquares model for Effec • The model was run sep of yield and purity by RP • The fractional factorial m inclusion of all single fac interactions as model efinteractions as model ef resource and time const was not possible. TECH LOGICBIO TECH LOGICBIO ®BioTechLogic, Inc. ta analysis ically significant effects of factors uantity, the JMP statistical used applying the Standard Least ct Leveragect Leverage. arately for two output parameters P-HPLC. model was initially run with the ctor and some two-factor ffects (Resolution IV) Due toffects (Resolution IV). Due to traints, a higher resolution study Slide 10 Company Confidential
- 11. Statistical Model P Step Yieldp • The final model used for effects and two factor int • There were two significa i ifi t i t ti Tsignificant interaction. T graphically represented i plots and interaction conp TECH LOGICBIO TECH LOGICBIO ®BioTechLogic, Inc. Parameter Estimates for assessment all four primaryp y teractions listed above. nt main effects and one h l ihese conclusions were in the main effects leverage tour plots.p Slide 11 Company Confidential
- 12. Main Effect Lever a. UV at Start Collection c. Gradient Slope TECH LOGICBIO TECH LOGICBIO ®BioTechLogic, Inc. rage Plots for Step Yield b. UV at End Collection d. Resin Load Slide 12 Company Confidential
- 13. Statistical Model P Purity by RP-HPLy y • The final model used for primary effects (excludin two-factor interactionstwo factor interactions. TECH LOGICBIO TECH LOGICBIO ®BioTechLogic, Inc. Parameter Estimates for C r assessment included all four ng resin load) as well as three Slide 13 Company Confidential
- 14. Two-Factor Intera Purity by RP-HPLy y Step Yield (%) Gradient Slope (%/CV) 0 UV @ End (%) a. UV at End Collection x Gradient Slope TECH LOGICBIO TECH LOGICBIO ®BioTechLogic, Inc. action Contour Plots for C Step Yield (%) Resin Load (g/L resin) 0 UV @ Start (AU) b. UV at Start Collection x Gradient Slope Slide 14 Company Confidential
- 15. Confirmation • There is one area of th represents process failp p yield, run 8. In order to acceptance criterion is operated within its defioperated within its defi modified and two addit augment the initial des • We chose to tighten th as it was determined to significant primary effesignificant primary effe analysis. • None of the experimenp failed the acceptance c confirms the predicted study TECH LOGICBIO TECH LOGICBIO ®BioTechLogic, Inc. study. of Results he tested design space that lure space with regard to stepp g p o ensure that the step yield always met when the process is ned PARs the design space wasned PARs, the design space was tional runs were performed to sign. e gradient slope upper limit to 11% o be easy to control and the most ect for step yield in our initialect for step yield in our initial ntal runs for the tightened limitsg criterion for step yield. This design space from the initial Slide 15 Company Confidential
- 16. Design Space SpaceSpace TECH LOGICBIO TECH LOGICBIO ®BioTechLogic, Inc. e vs Defined Control Based on the models generated for h f th d fi deach of the defined process outputs, a three-dimensional plot was generated to graphically show the process control space with d h i llregard to the experimentally defined design space Slide 16 Company Confidential
- 17. Conclusions • The results of this DO design space from the • Based on our findings h f th thwe chose for the three appropriate with the ex • As a result we tighteneAs a result we tightene • The IPC limits for resin set appropriately. TECH LOGICBIO TECH LOGICBIO ®BioTechLogic, Inc. E study confirm the predictedy p e initial study. we determined that the IPC limits i t te input parameters were xception of Gradient Slope. ed our control limitsed our control limits. n load were also determined to be Slide 17 Company Confidential
- 18. Final Paramet Normal Ope Operating Parameter Normal Ope (NO UV at the beginning of pooling, % from peak max Target: >p g p UV at the end of pooling, % from peak max Targe Gradient slope, % per CV TargeGradient slope, % per CV Targe Performance Parameter In Resin Load, g/L * Note: The claimed experimentally confirmed range is slightly tighter than th support the higher end of the range. TECH LOGICBIO TECH LOGICBIO ®BioTechLogic, Inc. ter Limits rating Range Experimentally Confirmedrating Range OR) Experimentally Confirmed Ranges > 0.5 AU 0.4 – 0.6 AU t: 20% 10 – 30% et: 8% 4 – 11%*et: 8% 4 11% n-Process Acceptance Criterion 15 – 25 g/L he design space in that it takes the worst-case data point to Slide 18 Company Confidential
- 19. Thank You BioTechLogic Inc serBioTechLogic Inc. ser biopharmaceutical ind with companies to mwith companies to m resource and comme TECH LOGICBIO TECH LOGICBIO ®BioTechLogic, Inc. rves therves the dustry by collaborating eet their developmenteet their development, ercialization needs. www.biotechlogic.com www.processvalidation.com Slide 19 Company Confidential