Biosafety in Biomanufacturing v6 updated.pdf

Biosafety – BioManufacturing Considerations Image generated from Bing AI Image Generator A/Prof Bernard Loo [email protected] Learning objectives Have knowledge of potential biosafety issues in biomanufacturing production Describe various types o...

Biosafety – BioManufacturing Considerations Image generated from Bing AI Image Generator A/Prof Bernard Loo [email protected] Learning objectives Have knowledge of potential biosafety issues in biomanufacturing production Describe various types of biohazards in biologics Be able to deploy various protective measures Understand the importance of control measures 2  Bioprocess/Biomanufacturing Bioprocessing (or biomanufacturing) refers to the production of biological materials such as vaccines, protein-based drugs, antibiotics or antibodies and cell-based drugs from living cells. Living cells such as yeast, bacteria, and animal cells are typically used. Insulin Herceptin (protein-based drug) (antibody) Chemical Pharmaceuticals Biopharmaceuticals MERCK/Schering-Plough Pharmaceuticals Amgen Zocor Epogen (Simvastatin) (Erythropoietin) v Size: 419 Dalton v Size: 30,400 Dalton v Decreases cholesterol synthesis v Treatment for anemia v Complex molecular structures v Simple chemical structures v Manufactured via biological processes v Manufactured via chemical processes (mammalian cells) Biological Products/Biopharmaceuticals Annual sales of approximately $343 billion in 2021 (https://doi.org/10.1038/s41587-022-01582-x) Includes vaccines, blood and blood comp., allergenics, gene therapy, recomb. therapeutic prot. Product Indications Factor VIII Hemophilia A Tissue plasminogen activator Myocardial infraction Erythropoietin Anemia Kymriah Acute lymphoblastic leukemia, large B cell lymphoma GLP-1 Type 2 diabetes  Insulin: Insulin: The First Biologic Drug The First Biopharmaceutical Product Discovery of Insulin Discovered by Frederick Banting in 1921 at the University of Toronto. Banting and Best treated diabetes using pancreatic extracts from dogs Dr Charles Sir. Frederick Best Banting 1899-1978 1891-1941 Dr James Dr John Collip Macleod 1892-1965 1876-1935 Discovery of Insulin In 1922, human clinical trials using insulin to treat diabetes was conducted with remarkable success Frederick Banting and J.J.R.Macleod were awarded the Nobel Prize in 1923 for their work on insulin Nobel Prize, 1923 Production of Insulin Sources of Insulin Pigs Cows Purified Insulin Recombinant Insulin In 1973, Herbert Boyer and Stanley Cohen showed that DNA from different species can be cut and pasted together This new DNA does not exist in nature Herbert Boyer A new genetic entity is created Called Recombinant DNA Technology This technology formed the basis for the first biopharmaceutical company Genentech Stanley Cohen  Production of Recombinant Human Insulin Bioreactor Human insulin gene The bacteria multiply and produce large quantities of human insulin in a bioreactor E. coli bacteria Bacteria are removed by filtration and sterilization, leaving behind the insulin DNA encoding for insulin is inserted into E.coli bacteria Further purification yields ultra pure crystals of human insulin Process Flowsheet of Human Insulin 12 Development of. Sus. Bioprocess ISSUES Based on the insulin example, what are the potential issues of obtaining insulin from animals sources and recombinant sources? 13 Biosafety Importance Eye drops contaminated with Pseudomonas aeruginosa 80 infected>, 4 deaths, 14 loss of vision https://edition.cnn.com/2023/05/19/health/ezricare-eye-drops-recall-update/index.html 14 Incidents related to Products of Biologic Origin 15 Incidents related to Products of Biologic Origin 16  Virus contamination in BioPharma Industry There have been several reported cases of viral contamination in large-scale cell culture processes Ø MMV (mouse minute virus) Ø Reo-3 Ø Calicivirus Ø Porcine circovirus Although rare, these events can potentially have significant consequences Ø Product impact Ø Long term facility shutdowns for decontamination/reboot Ø Disruption of medicine supply to patients Ø Business impact Adapted slides - Courtesy of Dr Yusdy Phan, Amgen 17  Typical Hazards in Biologics Guidelines for Process Safety in Bioprocess Manufacturing Facilities 18 by Center for Chemical Process Safety Additional considerations for biosafety 1. Procedural and facility factors 2. Equipment 3. Host Cells 4. Raw Materials 5. In-Process Control Testing 19 1. Procedural and facility factors: Air control HVAC, Airflow: Directional air, pressure gradients, airbreaks, separation of manufacturing area from offices, airlocks* 20 1. Procedural and facility factors: Biomanuf Room Design and Operations Facility walls, ceilings, lights: epoxy coated-hard plastic, welded vinyl sheet systems, sealable, easy to clean (http://www.daldrop.com/) Facility sanitation: Decontamination (wash hands, sticky mat and gowning), housekeeping, routine cleaning and disinfection, pest and rodent control Medical Surveillance Monitoring Biosafety Level Classifications Globally 22 Clean Room standards - US ISO 14644-1 (US FDA) ISO US FED STD Max particles/ft3 Equivalent 209E (rest/ops) > 0.1um > 0.2um > 0.3um > 0.5um > 5um Class 1 35 7 3 1 ISO Class 3 Class 10 350 75 30 10 ISO Class 4 Class 100 750 300 100 ISO Class 5 Class 1,000 1,000 7 ISO Class 6 Class 10,000 10,000 70 ISO Class 7 Class 100,000 700 ISO Class 8 100,000 23 Clean Room Standards – EU & ISO 14644-1 Max particles /m3 Equipped Equipped Equipped Equipped conditions conditions conditions conditions ISO EU (rest) (rest) (ops) (ops) Equivalent 0.5 um 5 um 0.5um 5um Class A 3500 0 3500 0 ISO Class 5 Class B 3500 0 3500000 2000 ISO Class 5 (rest) ISO Class 7 (ops) Class C 350000 2000 3500000 20000 ISO Class 7 (rest) ISO Class 8 (ops) Class D 3500000 20000 na na ISO Class 8 (rest) 24 Conventional Room Classifications for Biologics Area Classification (ISO) Classification (EU) Fermentation/Cell Culture 7 (rest), 8 (ops) or 8 (rest) C or D Buffer/Solution Preparation 8 (rest) D Purification 7 (rest), 8 (ops) C Bulk Filling 5 (rest & ops) A 25 1. Procedural and Facility Factors: Material and Personnel Flow 26  1. Procedural and Facility Factors: Environmental Monitoring Microbial Monitoring and Control Sample air, surfaces, water, and personnel to detect the presence of microorganisms such as bacteria, molds, and yeasts. Methods includes agar plate methods, impaction air sampling, settle plates, and swab sampling. Control: H2O2, Superchlor, Bleach 27 1. Procedural and Facility Factors: Waste Handling Used cell culture media, fermentation Biokill tank, Autoclave Assess: pH, harmful chemicals, biological demand, solids 28 2. Equipment – Sterile transfers and containment Closed systems Aseptic transfer Aspetic Port – Nova Septum 29 2. Equipment – Maintaining sterility and materials CIP & SIP “Cleanable”: electropolished 304 and 304L, 316 and 316L, 316SS Gasket Polymers: FPM, EPDM, PTFE, Silicone 30 Types of metal Chemical Eng Design 31 Piping & Valves L/D ratio = 2 maximum Valves 32 2. Equipment – Transferring aseptically and decontamination Autoclaves BSCs 33 3. Host cells: Cell Banking Master and working cell bank under lock and key. Potential biosecurity risk/contamination Sometimes MCB is potential source of contamination: Cells are checked for endogenous and exogenous microbe contamination 34 3. Host Cells: Approved Cell Lines for Pharma (Biologics) E.coli Saccharomyces cerevisiae Pichia Pastoris CHO BHK Murine myeloma Hybridomas HEK293 HT-1080 35 4. Raw Materials: Cell Culture Media Main Components: ØWater ØLow molecular weight nutrients ØNon-nutritive components ØHigh molecular weight factors Ideally it can be serum-free and chemically defined ØNon-animal sources ØLess lot to lot variability 36 5. In-Process Control Testing: Virus Assays Virus Assays Typically Used in Industry *Orthogonal testing and replicates are required due to inherent limitation of quantitative virus assays − In vitro viral assay − In vivo viral assay − Transmission electron microscopy (TEM) − Mouse Antibody Production (MAP) − Hamster Antibody Production (HAP) − qPCR 37 But don’t forget: Computer Security 38  Case Study/Discussions Growing interest in open ballroom manufacturing systems with functionally closed systems VERSUS conventional manufacturing plant design What would be the biosafety concerns if we have: HEK-293 perfusion plant process expressing a recombinant protein and a downstream post-viral filtration unit purifying (CHO Koenigsberg, Andrea L., et al. "MIT CAACB Risk Assessment Case Study: Assessing virus cross-contamination risk between two simultaneous processes in recombinant protein)? an open biomanufacturing facility." PDA Journal of Pharmaceutical Science and Technology 77.2 (2023): 115-132. (Adapted) Risk Assessment Quality Risk management workflow from ICH (adapted) Risk Question What is the risk of a viral contaminant escaping the HEK-293 perfusion bioreactor? Using FMEA: doi:10.5731/pdajpst.2021.012691 Risk Reduction Example of risk reduction steps doi:10.5731/pdajpst.2021.012691  Process Step with acceptable risk levels No risk reduction needed 43