dipesh-shah-catalent-pharma-solutions-usa.ppt
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- 3. Compatibility Assessment of a Model Monoclonal Antibody Formulation in Glass and in Blow-Fill-Seal (BFS) Plastic Vial Delivery Formats Dipesh Shah, Ph.D. Senior Development Engineer Advanced Delivery Technologies, Woodstock, Illinois ©2014 Catalent Pharma Solutions. All rights reserved
- 4. Presentation Outline 1. Advanced Aseptic Delivery Technology: Blow- Fill-Seal 2. General Requirements for Container Qualification and examples of Protein-Container Interactions 3. Case – Study: Compatibility assessment of a model monoclonal antibody formulation with AdvaseptTM and Glass vial
- 6. Quality/Sterility Assurance Minimizing Particulates Stability/ Compatibility Reduce risk factors for sterility challenges through automation Elimination of glass particles and delamination with significant reduction in foreign particulates Medical grade polypropylene resin for excellent chemical and physical properties Advanced Aseptic Filling Technology Why Glass Free? Safety/Reduced Product Loss Plastic construction reduces risk of breakage and simplifies opening Advanced Delivery Technologies for Parenterals to Mitigate Risk Associated with Aseptic Processing
- 7. Minimize Risk: Simplify the Process, Reduce Variables In just 15 seconds, the container is formed, filled and sealed in ISO 5 aseptic conditions
- 8. Minimizing Variables and the Footprint in Aseptic Manufacturing Traditional Glass Vial Filling Vials Stoppers Blow Fill Seal Sterilized Stopper insertion for vials Form Fill Insertion Seal Prequalified Resin Downstream equipment Utilizing the automated aseptic design of BFS, the technology eliminates traditional manufacturing steps, reduces the required controlled space and decreases the risk of contamination associated with traditional aseptic practices Glass Vial Filling Large controlled space Glass management and process control Blow Fill Seal Class A space 10 sq. ft Sterile container is formed at the time of fill
- 9. Leveraging Advanced Aseptic Process Blow Fill Seal Technology Reference: Verjans, B. Reed, C. (2012). “Assessing Filling Technologies for Contamination Risk.” Biopharm International. 25(3), pp. 46-58. The BFS process minimizes the risk of contamination by reducing particles, process steps & human interaction Potential risk of contamination by filling technology based on air quality and exposure time
- 10. 2: General Requirements for Container Qualification and Examples of Protein- Container Interactions
- 11. General Requirements for Container Qualification Safety – • Are we maintaining sterility? • What are we leaching? Efficacy – • Are we adequately protecting the dose? • Is there binding -- adsorption/absorption? • Are we delivering the dose?
- 12. Container Interactions with Protein Therapeutics Discussion Topics 1.What are the product development concerns for container interactions with protein products? 2.What testing is needed to understand container interactions with protein therapeutics?
- 13. Liquid Formulation Containers (Injectables)– Potential Materials Enhanced System Container Materials Closure Materials Vials or Ampoules (SVP) Glass/AdvaseptTM Vials with Butyl rubber/Teflon coated butyl rubber Prefilled syringes with plungers Glass or Plastic Staked needle or luer lock (plastic or rubber) Butyl rubber Teflon coated butyl rubber IV Infusion (LVP) Glass/Flexible Bags with spike ports/AdvaseptTM Polyethylene
- 14. General Concerns for Protein-Container Interactions Container Concerns Product concerns • Protein adsorption • Protein loss • Headspace • Agitation causes protein aggregation (AdvaseptTM Advantage) • Organic leachables (eg. Silicone, DEHP) • Toxicity of leachable • Protein aggregation/Immunogenicity • *Trace metals (eg. Iron, Tungsten, Aluminum) • Eg. Iron – Iron Leachable catalyzed protein degradation reactions • Eg. Tungsten – Protein Oxidation (AdvaseptTM Advantage) • Eg. Aluminum – Aluminum Phosphate resulted in visible particles (AdvaseptTM Advantage) *www.pqri.org/workshops/podp11/pdfs/markovic.pdf , “Regulatory perspective safety qualification of extractable and leachable,” Feb. 22-23, 2011, Ingrid Markovic, Ph.D.
- 15. Challenges to fill Biologics with Blow-Fill- Seal (BFS) Challenges: 1. Impact of elevated temperature of the molded plastic during filling on the stability of the biologic formulation 2. Impact of gas permeation from semi- permeable BFS plastic container on degradation of biologics (for eg. Oxidation) 3. Impact of potential leachables from BFS processed plastic container system on biologic stability and safety of the formulation.
- 16. Temperature Profile of Resin and Container
- 17. Temperature Profile of solution Conclusion: The molded plastic process was optimized by Catalent to reduce the temperature of the solution (0.5 mL fill) close to 40°C at time of fill and the assumption is that the temperature of the solution would fall steadily after the units come out of the fill suite and packaged under ambient conditions.
- 18. Oxygen and Carbon Dioxide Gas Permeation Data Conclusion: Carbon Dioxide has higher permeation rate relative to Oxygen because Carbon Dioxide partitions into the container to a greater extent relative to Oxygen. 0.0E+00 2.5E-02 5.0E-02 7.5E-02 1.0E-01 0 10 20 30 40 cc*mm/(cm 2 *day) Temperature (C) Gas Permeation Rate vs Temperature (Normalized) Oxygen Carbon Dioxide
- 19. Water- Vapor Transmission Rates 0.0E+00 1.0E-05 2.0E-05 3.0E-05 4.0E-05 5.0E-05 5 10 15 20 25 30 35 40 g*mm/(mm 2 *day) Temperature (C) Water Vapor rate (normalized) vs Temperature (C) Conclusion: Higher the Temperature, greater the water vapor permeation rates
- 20. Case – Study: Compatibility assessment of a model monoclonal antibody formulation with AdvaseptTM and Glass container system
- 21. Monoclonal Antibody Formulation Recipe + Preparation . Table 1: Model mAb Formulation Components Amount (mg/mL) Model mAb ( Mol. Wt. 144 kDa) 10 Polysorbate 80 0.7 Sodium Citrate 6.5 Sodium Chloride 9.0 pH 6.5 The monoclonal antibody formulation was filtered with a 0.2 micron Nalgene filter unit and filled in glass vials into which uncoated stoppers were placed. The mAb formulation was filtered into a sterile bag and shipped to Blow-Fill-Seal (BFS) manufacturing facility. The formulation was aseptically transferred in BFS AdvaseptTM stoppered vials.
- 22. mAb Compatibility Assessments (Glass/Advasept Vial formats) Parameter Method Target Range pH USP 791 6.5 + 0.3 Appearance Visual Inspection Report Results Potency UV (280 nm) T=0 + 10% Activity Report EC50 Stability Size-Exclusion Chromatography (SEC) Report % Monomer and % High and low molecular weights Nanoparticle Tracking Analysis (NTA)5 Report sub-micron particle size analysis Sodium Dodecyl Sulfate- Polyacrylamide Gel Electrophoresis (SDS-PAGE) Report % Area (HC +LC) Capillary Isoelectric focusing (cIEF) pI (% of each peak) Peptide Mapping2 % Chemical Modification Leachables3 Polar Leachables (HPLC-UV) Report Irganox 1010 levels Semi-Volatile Leachables (GC-MS) Report Aromatic Hydrocarbon levels Volatile Leachables GC-FID Report Volatile Leachable levels4 Metals (ICP-MS) Report all metals except Na, I Bacterial Endotoxin1 USP 85 Report Results Bioburden1 USP 61 <10 CFU/mL 1 only performed at 0 time-interval 2 Only performed at 4 month time-interval 3 Only performed at 9M time-interval 4 Isopropyl alcohol, Methyl Ethyl Ketone, Trichloroethylene, Hexane, 2-methyl pentane, 3-methyl pentane, Methylcyclopentane, Cyclohexane and Heptane 5 Performed on 12 M samples
- 23. Results 1 -Potency Potency: a) UV: The UV data indicates no apparent change in potency for both vial formats upon pre and post-fill and upon storage to 9 months at 5°C were within target range. b) Complement Dependent Cytotoxic Assay: The potency was determined using responsive cell line in a complement dependent cytotoxic assay using a fluorescence read out. Comparison of a dilution series with standard, formulation in glass and the AdvaseptTM vial were generated (Figure 1). Figure 1: Dose Response Curve for model mAb formulation x axis 0.001 0.01 0.1 1 10 0 1000 2000 3000 Graph#1 y = ( (A - D)/(1 + (x/C)^B ) ) + D: A B C D R^2 Advasept 2 (Advasept 2: Conc vs MeanValue) 3574.447 1.618 0.373 247.764 0.996 Glass vial 2 (Glass Vial 2: Conc vs MeanValue) 3511.099 1.729 0.482 325.305 0.998 Advasept 1 (Advasept 1: Conc vs MeanValue) 3534.978 1.711 0.391 320.817 0.998 Glass vial 1 (Glass vial 1: Conc vs MeanValue) 3568.287 1.728 0.46 338.779 0.999 Conclusions: The activity data shows comparable potency values between the Glass and AdvaseptTM vials upon storage to 24M/5°C (All data not shown).
- 24. Result 3 -Nanoparticle Tracking Analysis (NTA) Particles AdvaseptTM Glass Vial D50 126 + 17.5 94 + 10.1 nm D90 205 + 35 nm 144 + 13.4 nm Total Concentration (particles/frame) 13.95 + 3.2 13.47 + 3.0 Total Concentration (particles/mL) 2.83e8 + 6.5e7 2.74e8 + 6.1e7 Mean sub-micron particle count data (Nanoparticle Tracking Analysis) for Monoclonal antibody formulations stored in Glass and AdvaseptTM vials for 12M/5°C. Conclusions: Slightly higher size mode of aggregates were observed in AdvaseptTM than in the glass vials, however, the total number of particles are statistically comparable between the two container closure systems.
- 25. Result 2 SEC-UV Aggregation Data Conclusion: Stability data indicates higher levels of high molecular weight species in Glass relative to AdvaseptTM vial. 0 1 2 3 4 5 6 0 6 12 18 24 % High Molecular Weight Species Months SEC-UV - Aggregation data (5C)
- 26. Results 4-Peptide Mapping Results Type Sample % Modification Oxidation Glass Vial 10.8 AdvaseptTM Vial 4.5 Deamidation Glass Vial 7.2 AdvaseptTM Vial 7.6 Conclusions: The oxidation levels were higher in Glass relative to AdvaseptTM vials and could be attributed to mAb’s surface interaction with glass surface causing more oxidation than in plastic AdvaseptTM vials. Procedure: Aged Glass and AdvaseptTM vials (4M/5°C) were subjected to peptide mapping by initially denaturing and reduction of the mAb with DTT, alkylation with Iodoacetamide, followed by clean-up on a column, and digestion with Trypsin and ASP-N. The peptides were separated on a UPLC column and UV and MS detectors were employed. (Chromatograms provided in the appendix section).
- 27. Results 5 (pH, Appearance, cIEF and SDS-PAGE) All other stability test results for parameters listed below were comparable between Glass and AdvaseptTM vial formats. 1. pH 2. Appearance 3. cIEF 4. SDS-PAGE Bioburden results performed at t=0 for both vial configurations were below <10 CFU/mL.
- 28. Results 6 (Leachable Data Analysis) The monoclonal antibody formulation samples stored for 9M at 5°C were analyzed for various leachables listed below: 1.Volatile: GC-FID 2.Semi-Volatile: GC-MS 3.Polar: HPLC-UV 4.Metal leachables: ICP-MS Results: The leachable data indicates comparable leachable profile for monoclonal antibody formulations stored in Glass and AdvaseptTM vials except for Isopropyl alcohol. The Isopropyl alcohol content was determined to be higher in Glass vial (14 mg/mL) relative to AdvaseptTM vial (1.2 mg/mL) and was attributed to cleaning procedure used for glass vials prior filling of the monoclonal antibody formulation.
- 29. Conclusions 1. This study confirms compatibility of monoclonal antibody formulation in glass and AdvaseptTM vial demonstrating plastic BFS vial suitability for protein therapeutics. 2. Stability data indicates higher levels of high molecular weight species in Glass relative to AdvaseptTM vial. 3. Affinity data indicates comparable potency levels in Glass and AdvaseptTM vial. 4. Leachable data indicates comparable leachable profiles in Glass and AdvaseptTM vial. 5. No significant differences were identified between Glass and AdvaseptTM vials over the 9 months analyzed to date.
- 30. ACKNOWLEDGEMENTS Catalent- Woodstock Catalent – Kansas City 1. Waiken Wong, Ph.D. 1. Vincy Abraham, Ph.D. 2. Bill Hartzel 3. Natasha Hults Catalent - RTP 4. Kay Schmidt 1. Thomas Luntz 2. Courtney Jones Catalent – Madison 3. James Mclean 1. Gregory Bleck, Ph.D. 4. Vicki Wards 2. Ian J. Collins, Ph.D. 3. Process Development Team
- 31. QUESTIONS ???
- 32. APPENDIX
- 33. cIEF Analysis cIEF Chromatogram Time 0 a) Glass Vial b) AdvaseptTM vial Magnified Chromatograms Time 0 a) Glass Vial b) AdvaseptTM vial The chromatograms showed no comparable charge distribution between the Glass and AdvaseptTM vial.
- 34. Impact of Elevated Temperature of the Molded Plastic during filling on the stability of biologic formulation T=0 data Parameters Bulk Solution Glass Vial AdvaseptTM Vial pH 6.4 6.6 6.6 Potency (UV) (mg/mL) 10.8 10.8 10.9 SEC 2.2% High Mol. Wt. 97.8% Monomer 2.1% High Mol. Wt 97.9% Monomer 2.1% High Mol. Wt. 97.9% Monomer SDS Similar Profile (Reduced/Non Reduced) Figures – Appendix section cIEF Major PIs (%)* pI Range 9.38-9.65 (7 peaks) Major PIs (%) pI Range 9.42-9.74 (8 peaks) Major PIs (%) pI Range 9.42-9.74 (8 peaks) Conclusion: Data from stability indicating parameters between the bulk mAb, glass and AdvseptTM vial drug product indicates that the BFS process is amenable to Biologics. Peak pI % Area 1 9.38 11.0 2 9.40 14.0 3 9.45 11.9 4 9.50 34.0 5 9.56 3.49 6 9.59 20.7 7 9.65 5.0 Peak pI % Area 1 9.42 15.9 2 9.44 3.2 3 9.49 18.8 4 9.54 31.8 5 9.58 6.2 6 9.62 18.9 7 9.67 5.1 8 9.74 0.1 Peak pI % Area 1 9.42 15.8 2 9.44 3.1 3 9.49 18.6 4 9.54 31.9 5 9.58 6.2 6 9.62 19.1 7 9.67 5.3 8 9.74 0.2 *analysis occurred at separate time using different cIEF cartridge which may explain slight differences and possible poor resolution of peak 8 which is only 0.1% of the total area. Peak profiles look similar. cIEF (peaks)
- 35. SDS – PAGE Analysis SDS-PAGE (Non-Reduced) Lane 5 – Glass Vial Lane 6 – AdvaseptTM Vial SDS-PAGE (Reduced) Lane 5 – Glass Vial Lane 6 – AdvaseptTM Vial
- 36. Magnified UPLC/UV Chromatograms of Trypsin Digested Monoclonal Antibody AU -0.02 0.00 0.02 0.04 0.06 0.08 0.10 0.12 0.14 0.16 0.18 0.20 0.22 0.24 0.26 0.28 0.30 0.32 0.34 Minutes 10.00 15.00 20.00 25.00 30.00 35.00 40.00 45.00 50.00 55.00 60.00 65.00 70.00 75.00 80.00 85.00 Glass Vial (Bottom Trace), AdvaseptTM Vial (Top Trace)
- 37. Magnified UPLC/UV Chromatograms of ASP-N Digested Monoclonal Antibody AU 0.00 0.02 0.04 0.06 0.08 0.10 0.12 0.14 0.16 0.18 0.20 0.22 0.24 0.26 Minutes 10.00 15.00 20.00 25.00 30.00 35.00 40.00 45.00 50.00 55.00 60.00 65.00 70.00 75.00 80.00 85.00 Glass Vial (Bottom Trace), AdvaseptTM Vial (Top Trace)
- 38. Let us meet again.. We welcome you all to our future conferences of OMICS International 2nd International Conference and Expo on Parenterals and Injectables On October 24-26, 2016 at Istanbul, Turkey http://parenterals- injectables.pharmaceuticalconferences.com/