QbD based Development and Characterization of a Cell Culture Process for Therapeutic Proteins

Transcription

1 QbD based Development and Characterization of a Cell Culture Process for Therapeutic Proteins Green Cross Corp. Green Cross Research Center Yong Jae Kim (YJKIM@greencross.com) Biologics World Korea 2015

2 Agenda Company Introduction QbD and Cell Culture Process Development Case Studies

3 Green Cross History 1967 Established as Sudo Microorganism Medical Supplies Co Production of the Nation s First Plasma Fractionation Product Hepatitis B Vaccine for the 3rd Time in the World (Hepavax) World s 1st Epidemic Hemorrhagic Fever Vaccine (Hantavax) 1993 World s 2nd Varicella Vaccine (Suduvax) 2009 Construction of Ochang Plant for Bioproducts 2009 Construction of Hwasun Plant for Vaccine Products 2009 Approval of Seasonal Influenza Vaccine (GC Flu) H1N1 Influenza Vaccine (GreenFlu) 2010 Approval of Recombinant Human Factor VIII (GreenGene F) 2011 Approval of Shinbaro (Herbal Drug) 2012 Approval of Hunterase (Hunter Syndrome) 2013 Construction of R&D Center

4 Sales Trends Profile Summary Sales $950M Established 1967 $807M $887M IPO(KOSPI) 1978 Employees Market Capital 1,500 $1.37B $71.6M Operating Profit 2013 $84.6M 2014 $85.4M 2015(E)

5 Investment in R&D Sales Composition R&D Spending $91M Rx 19.4% Others 16.5% Vx 31.6% PD 32.5% R&D Spending as a % of Sales $71M 8.8% $80M 9.0% 9.6% (E)

6 Facility Overview Head Office and R&D Center Eumseong Plant Seoul Yongin - OTC Hwasun Plant Ochang Plant - Vaccine (FLU and Others) - Plasma Fractionation - Recombinant

7 R&D Capability Plasma Fractionation Vaccine Antibody Engineering Recombinant Protein

8 What is Quality by Design? Risk Assessment DOE QbD Process Understanding QRM Design Space Statistics MVDA Systematic Approach

9 Overall QbD Approach FDA s GMPs for the 21 st Century and the PAT initiative QbD related guidelines ICH Q8, 9, 10, 11 European Journal of Pharmaceutics and Biopharmaceutics 81 (2012)

10 Systematic approach to Applying QbD Pharmaceutical Quality by Design(QbD) Principles SeerPharma Pty Ltd

11 Upstream Process Development Advances in The Development of Biopharmaceuticals, Invitrogen

12 Upstream Manufacturing Process Trends in Biotechnology Vol.28 No.5

13 Cell Culture Process Operating Parameters Affect Process Performance & Product Quality mabs 2:5, ; September/October 2010; 2010 Landes Bioscience

14 Identification of Operational Parameters Potential Critical Process Parameters in Cell Culture Production Temperature ph Agitation Dissolved oxygen Medium constituents Feed type and rate Howard L. Levine, Ph.D. BioProcess Technology Consultants, Inc. FIP Quality International 2007 Conference

15 Combination of Risk Assessment & Statistical Design of Experiments (DoE)

16 Process Development and Characterization Case I : Flask Inoculum Expansion Project: Therapeutic Proteins I Cell line: CHO-DG44 Medium: Commercial Medium Culture vessel: Shake Flask Culture volume: 100mL Incubator: CO 2 Incubator and Orbital Shaker

17 Operational Parameters Analysis of the Flask Inoculum Expansion Failure Mode Effect Analysis (FMEA) - Severity of the excursion: S - Occurrence of the excursion: O - Detection of the excursion: D - Risk Priority Number(RPN) = S X O X D

18 Design of Experiments Run Initial VCD Temp. CO 2 Duration (E5 cells/ml) ( ) (%) (hr) Remarks Resolution IV 4 center points Augmentation Axial points

19 Prediction Profiler

20 Prediction Formulas Response Polynomial equation of the response in terms of coded factors Statistical significance R 2 R 2 Adj p-value VCD CP*+3.23A-6.09B+2.39C+4.41D-0.58AB-4.32B C BD-4.61D < Viability CP*-10.57B+5.56C-2.2D-7.96B BC-2.19BD-4.11D < A: Initial VCD, B: Temp., C: CO 2, D: Duration CP*: prediction of the response at the Center Point

21 Design Space (Contour Plots) *NOR: Normal Operational Range

22 Process Development and Characterization Case II : Production Bioreactor Project: Therapeutic Proteins II Cell line: CHO-DG44 Medium: In-house medium Culture system: Glass type STR Culture volume: 2.5 L

23 Operational Parameters Analysis of the Production Bioreactor Failure Mode Effect Analysis (FMEA) - Severity of the excursion: S - Occurrence of the excursion: O - Detection of the excursion: D - Risk Priority Number(RPN) = S X O X D

24 Design of Experiments Run Temp. ph Timing of Prod. shift ( ) (E5 cells/ml)

25 Prediction Profiler

26 Regression models for the production bioreactor responses Response Polynomial equation of the response in terms of coded factors Statistical significance R 2 R 2 Adj p-value Yield CP*-416A+156B+662C-439AB-327AC-435BC 901A ABC Impurities#1 CP*+1.07A+1.59B+6.04C+3.25AC+1.61BC-8.79C Impurities#2 CP*+7207A+5040B+2401C+3425AB+967AC+700BC+5116C QA#1 CP*+3.72A+5.90B+1.97C-2.00AB+1.54AC 0.77BC 8.62B ABC QA#2 CP*+11.91A+7.92B+1.61C 0.88AB+1.55AC-0.02BC-13.39A ABC QA#3 CP*+8.25A+5.99B 2.11AB 12.54A QA#4 CP*+1.1A+1.13B+0.15C 0.19AB+0.33AC+0.24BC 2.25B ABC QA#5 CP*+8.68A+4.83B+0.41C+0.44AB 1.52AC+3.36BC 9.07A ABC A: Production Temp., B: Production ph, C: Timing of production shift CP*: prediction of the response at the Center Point

27 Design Space (Contour Plots) NOR NOR NOR X : ph, Y : Temp. X : Prod. shift timing, Y : ph X : Prod. shift timing, Y : Temp. *NOR: Normal Operational Range

28 Summary The concept of QbD aims at gaining a deeper knowledge of the product and the manufacturing process starting with product development. Case studies describe the application of QbD principles to the flask inoculum expansion step and the production bioreactor step of therapeutic protein, which resulted in the definition of a design space for the cell culture process. Characterization studies were performed using a multi-fractional DoE approach to determine the impact of the identified potential CPPs, both individually and as interactions, on CQAs The design space was set in such a way to ensure consistent delivery of a cell culture process for the requirements set for the therapeutic proteins.

29 Thank You.