Introduction to Bioprocessing Sample Slides
- 1. Introduction to Bioprocessing Cambridge Healthtech Institute Peptalk Palm Springs, CA Presented by Susan Dana Jones and Sheila Magil BioProcess Technology Consultants www.bptc.com
- 2. BioProcess Technology Consultants World leader in providing strategic, technical, regulatory, and business consulting services to biopharmaceutical industry Founded in 1994; 20+ year proven track record Team of 13 consultants has provided consulting services to over 500 clients across six continents Process Development Quality & Regulatory Manufacturing Strategy Supply Chain Program Management Due Diligence 2 From Clone to Clinic
- 3. Instructors Sheila Magil, Ph.D. Senior Consultant, BPTC Sheila has over 20 years of experience in quality and analytical method development for biologics, peptides and small molecules. Her expertise includes quality assurance, protein and peptide biochemistry, and analytical development. Sheila has implemented quality systems and has managed external analytical and QC activities for multiple biopharmaceutical products. 3 Susan Dana Jones, Ph.D. Vice President and Senior Consultant, BPTC Susan is a seasoned biotechnology entrepreneur with experience in product development, outsourcing, and strategic planning. She is a subject matter expert in cell line development and characterization for biosimilar, new biopharmaceutical, and vaccine development programs. She has broad knowledge of regulatory requirements for manufacturing products for human use and has prepared CMC sections of multiple regulatory submissions. From Clone to Clinic®
- 4. Introduction to Bioprocessing Course Content Definition of biopharmaceutical products Regulatory considerations in bioprocessing General timelines for biopharmaceutical product development Critical CMC activities: • Project management and tracking • Analytical methods • Expression systems • Downstream processing and viral clearance/inactivation • Formulation • Stability Cost considerations Process qualification Comparability 4 From Clone to Clinic®
- 5. What are Biopharmaceutical Products? Therapeutic proteins including antibodies, nucleic acids, gene therapy viral vectors • Generally produced by recombinant DNA technology Excluded from definition are traditional biologic products • Non‐recombinant vaccines, plasma‐derived proteins Focus of this training course is on production of biopharmaceutical products, especially monoclonal antibodies, in mammalian cell culture 5 From Clone to Clinic® Example Biopharmaceutical Product MW # of Amino Acids # of Protein Chains Glycosylated Insulin ~6 kDa 51 2 No Growth Hormone ~22 kDa 191 1 No Erythropoietin 30‐34 kDa 165 1 Yes Coagulation Factor IX ~52 kDa 461 1 Yes Monoclonal antibody (IgG1) ~150 kDa ~660 4 Yes Factor VIII ~240 kDa 2133 1 Yes
- 6. Project Management: Integration of Development Activities When to scale up, what to use for GLP tox, what scale, batch records From Clone to Clinic® Analytical methods: Understand the molecule Microbial or mammalian, novel or established, accelerated or standard? Expression System Platform or de novo, small scale systems, QbD, DOE, applied knowledge Process Development Scale up and Production Analytical Methods
- 7. Protein Structure Proteins have an inherent degree of structural heterogeneity Primary Structure • Amino acid sequence • Held together by peptide (amide) bonds Secondary Structure • Held in place by hydrogen bonding Tertiary Structure • Driven by hydrophobic and ionic interactions Quaternary Structure • Cluster of similar or different proteins Primary Structure Secondary Structure Tertiary Structure Quaternary Structure Amino Acids ‐Pleated Sheet ‐Helix 7 From Clone to Clinic®
- 8. Classes of Analytical Methods Classes of Methods Specific Methods Spectroscopy UV‐Vis, CD, FTIR, Fluorescence, Raman Chromatography Reverse Phase, Size Exclusion , Ion Exchange, Hydrophobic Interaction, Affinity Electrophoretic SDS‐PAGE and Normal Phase, Reduced and Non‐reduced, 2‐Dimensional, Isoelectric focusing, capillary Coomassie and Silver stains Immunophoretic Western Blot, ELISA Immunoassays Bioassays, ELISA Potency Bioassays, Cell‐based, Binding Microbiological Analyses Bioburden, Sterility, Viability, Microbial Contamination, Endotoxin Chemical (Compendial) USP/EP/BP/JP raw material testing Other Analytical Ultracentrifugation, Particulate Matter, pH, Visual appearance 8 From Clone to Clinic®
- 9. Role of Analytics With each step, analytical methods have become increasingly important and front‐end loaded “The QbD process design starts with an intensive characterization of the product through a large array of biochemical and biophysical analyses at normal and stressed conditions…” The QbD process design starts with an intensive characterization of the product through a large array of biochemical and biophysical analyses at normal and stressed conditions…” Banerjee A. BioPharm Intl 23(5): 26‐40. 9 Banerjee AF. rBoiomPha rCmloInntl e23 (t5o): 2C6l‐4in0.ic® Process defines Product Well Characterized Biologics Quality by Design and Process Analytical Technologies Increasing process and product understanding 1980’s 2010’s
- 10. General Scheme for Biopharmaceutical Manufacturing Working Cell Bank (WCB) Inoculum Preparation Production Bioreactor Primary Recovery Purification Polishing Formulation Bulk Drug Substance (API) 10 From Clone to Clinic®
- 11. Common Production Hosts Critical issues to address Technical • Overproduction may cause aggregation or degradation by host, toxicity to host cell, inaccurate or incomplete processing • Post translational modification (glycosylation) Intellectual Property Rights Manufacturing Capabilities for Clinical and Commercial Supplies Bacteria • E. coli • Pseudomonas fluorescens Yeast • Saccharomyces cerevisiae • Pichia pastoris Mammalian cells • Chinese hamster ovary (CHO) • BHK and HEK (specific indications) • Per.C6 and other human lines 11 From Clone to Clinic®
- 12. An important trend – single use bioreactors Xcellerex XDRTM Bioreactor Sartorius Stedim Biostat® Culti‐bag Thermo Fisher (Hyclone) Single‐use Bioreactor ATMI NucleoTM Bioreactor GE Healthcare Wave Bioreactor 12 From Clone to Clinic®
- 13. Downstream processing of biopharmaceuticals Downstream Process Bulk Viruses Pyrogens Cells Nucleic acids Viruses Cell culture medium Host cell proteins CIP Nucleic acids Leakage Protein Raw Material (rDNA) Pure Bulk Product (Therapeutic) Ligand Leakage Microorganisms Pyrogens 13 From Clone to Clinic®
- 14. Column sizing and optimization Balance cost of production (COP), production rate (g/hr) and productivity (g/hr/L support) • Trade‐off between capacity, throughput, and cost • Multiple cycles vs. a larger column? • How frequently should media be replaced? • Support equipment sizing and cost Cycling small column in series Multiple small columns in parallel Single large column 14 From Clone to Clinic®
- 15. Product Comparability Primary structure • Amino acid sequence Secondary structure • Three‐dimensional structure of the protein (alpha helices, beta sheets, loops/turns) Tertiary structure • 3‐D structure of protein through interaction of the secondary structures • Increased regulatory emphasis on this aspect of structure Quaternary structure • Describes the three‐dimensional arrangement of protein subunits 15 From Clone to Clinic®
- 16. Product Consistency Potency Structure Stability Safety Efficacy Product Control Through Process Control Cell line Media and Feeds Culture Time Column Loading Intermediate Hold Time Formulation Buffer QbD PAT From Clone to Clinic®
- 17. Thank You! Susan Dana Jones and Sheila Magil BioProcess Technology Consultants, Inc. 12 Gill Street, Suite 5450 Woburn, MA 01801