Continuous mab Purification Process: Design Features and Practical Considerations

Transcription

1 Continuous mab Purification Process: Design Features and Practical Considerations Joanna Pezzini CASSS DC Area Discussion Group Meeting December 7, 2017

2 Outline Continuous Process Overview Continuous Technologies PAT Opportunities Closed Processing 2

3 Continuous Process Overview Continuous Technologies PAT Opportunities Closed Processing 3

4 Batch Purification Process Overview Entire batch is pooled in product hold tank between each process step. ProA Capture Viral Inactivation Anion Exchange CEX Capture Viral Filtration UFDF & Formulation Protein A bind & elute chromatography ph adjustment in stirred tank low ph hold time in stirred tank for viral inactivation Flow through chromatography or membrane Cation exchange bind & elute chromatography Virus pre filter and virus filter in series Diafiltration Final Concentration Excipient spike Final filtration Bottle fill

5 Continuous Purification Process Overview Continuous flow through all steps for the duration of the process. Steps are separated by small product break tank. Multi-Column Chromatography (ProA) Flow-Through Low ph Viral Inactivation Filtration Train (Includes AEX and Viral) Multi-Column Chromatography (CEX) Single Pass UFDF Protein A bind & elute chromatography Inline ph adjustment Residence time in SEC column provides low ph hold time for viral inactivation Depth Filter Sterile Filter AEX Membrane Virus Pre-Filter Virus Filter Cation exchange bind & elute chromatography Diafiltration Final Concentration Inline excipient spike Final filtration Bottle fill 5

6 Scale by Volume Scale by Volume Benefits of Continuous Processing Improved productivity Flexible capacity management (scale by time in addition to volume) Decreased downstream cost of goods Amenable for labile proteins Facilitates adoption of single use technologies time ProA Capture Viral Inactivation Anion Exchange CEX Capture Virus Filter UFDF time ProA Capture Viral Inactivation Anion Exchange Virus Filter CEX Capture UFDF Scale by Time

7 Scale Up of Batch Upstream Process Proposed design applies to processes from 200 L to 20,000 L. 200 L Scale 2,000 L Scale 20,000L Scale Column Diameter 4.4 cm 10 cm 30 cm System Piping Inside Diameter 1/32 1/8 1/4 Break Vessel Volume 200 ml 2 L 20 L Eluate Tank Volume 5 L 50 L 500L Small Volumes Enable Single Use Containers 7

8 Outline Continuous Process Overview Continuous Technologies PAT Opportunities Closed Processing 8

9 Multi-Column Chromatography- an Enabling Technology Feed Wash Elution Benefits Continuous feed Increased resin utilization (g/l resin ) Improved productivity (g/l resin hr) > 50% resin savings > 20% buffer savings Comparison of chromatography systems for batch process to demonstrate improved efficiency. Single Column Operation Multi-Column Chromatography Column Size 60 cm diameter x 20 cm height 30 cm diameter x 10 cm height Column Vol. 56 L 28 L total Resin Cost $616,000 $308,000 Process Time 8.2 hours 6 hours 9 Productivity 20 g/l resin hr 60 g/l resin hr

10 MAU Novel Flow-Through Hold Design for Viral Inactivation Base Next Unit Op Product Acid Base Product Acid Traditional Titration/Hold Method Continuous Titration/ Residence Time Method Continuous flow design Easily scalable Inline titration leads to consistency Process range min SEC Column Transition TIME (MINUTES)

11 Concentration Novel SPDF Achieves 99.75% Buffer Exchange Single Pass Diafiltration (SPDF) designed to meet or exceed buffer exchange equivalent to a 6 diavolume traditional process (99.75% buffer exchange). Achieved with three sequential dilutions 7.5X Product concentrated 7.5X with SPTFF membrane prior to each dilution to minimize buffer use Final Concentration = Initial Concentration Dilution Factor # stages Feed Retentate 1 Diluent 1 Retentate 2 Diluent 2 Retentate 3 Diluent 3 (DF Product) 11

12 Outline Continuous Process Overview Continuous Technologies PAT Opportunities Closed Processing 12

13 Process Flow Diagram ph Control Pressure and Volume Control Pressure Control 13 SPTFF: Single-Pass Tangential Flow Filter

14 Control Plan Arduino 14

15 Control Plan Controlled Variables (CV): ph 1 ph 2 P1 M2 P2 P3 P4 P5 P6 M3 P7 P8 P9 P10 Viral Inactivation Filter Train Single-Pass Diafiltration 6 Manipulated Variables (MV): Pump 1 Pump 2 Pump 3 Arduino 1 Pump 4 Arduino 2 Pump 5 Pump 6 Arduino 3

16 PAT Opportunities for Continuous Can PAT and automation improve product quality and/or process efficiency? Current ph feedback to control low ph viral inactivation acid and base addition rates Pressure feedback to control flow rate during chromatography or membrane filtration Volume feedback to control break tank volume inbetween continuous steps Product concentration to monitor UFDF step Proposed Aggregate concentration for wash/elution buffer composition of CEX step HCP, DNA concentration for AEX membrane lifetime Yield to evaluate chromatography resin lifetime Charge variants to determine bioreactor conditions 16 Steady state operation achieved with continuous manufacturing provides time to measure and respond to changes in product quality attributes.

17 PAT Decision Tree Can PAT and automation improve product quality and/or process efficiency? yes How fast is feedback needed? no No need for PAT Focus on automated on-line sample collection and intime analysis for batch release <5 min 5-10 min > 10 min Is inline instrumentation available? On-Line At-Line Focus on automated on-line sample collection for at-line analysis Inline PAT yes no No PAT Available to Implement Bring to Industry Attention (BPOG, Suppliers, etc.) 17

18 What can be achieved? In-line Flow Pressure ph Conductivity UV Concentration (Density, IOR, A280) Spectroscopy (Raman, NIR, DLS, MALS, IOR) On-line Chromatography based Methods: SEC for aggregate, fragments IEX for charge variants Mass Spectrometry based methods for: Oxidation Deamidation Glycosylation Di-sulfide bond modifications At-Line Automated/Robotic sample collection, handling, preparation, and injection into the following instrumentation: Enzyme-linked immunosorbent assay (ELISA) for host cell protein (HCP) Polymerase Chain Reaction (PCR) for deoxyribonucleic acid (DNA) 18 Routinely used in GMP biologics manufacturing facilities Not routinely used in GMP biologics manufacturing facilities

19 Automated Sampling Need for Continuous High sampling frequency, number of measurements, and number of sample points requires automated on-line sample collection and stream-lined off-line sample analysis 1 per day x 5 sample points x 10 measurements = 50 samples per day High sampling frequency to enable feedback to control process, monitor for drift in process over time Close location of sample points enables automated sample collection with single piece of equipment (sample lines feed to different sample points) 19

20 Outline Continuous Process Overview Continuous Technologies PAT Opportunities Closed Processing 20

21 Bioburden control Continuous process operated over several days or weeks must maintain low bioburden levels, similar to batch process requirements. Our approach for bioburden control: Gamma irradiated vessels and sanitization solution through chromatography/filter systems upon startup and as needed. 0.2μm filter Chromatography/Filter System 0.2μm filter 250 ml sterile bottle Sanitization Solution 250 ml sterile bottle 21

22 Bioburden Control- Vessels Vessels gamma irradiated. Vessels include 3-way valve on each of three tubing inlets. 1. Upstream inlet 2. Downstream outlet 3. Buffer addition line Vessels also include a vent filter and a stir bar. Vessels placed on stir plate that rests on scale. 22

23 Questions??? MedImmune s First Fully Integrated Continuous Pilot Scale Demo 30 L Brx + ATF 2.5 g/hr ProA Multicolumn Chromatography 1.7 g/hr POD AEX VF 11 g/hr Viral Inactivation 11 g/hr Diafiltration and Final Concentration 11 g/hr 23

24 Acknowledgements Bioprocess Engineering Purification Process Sciences Cell Culture & Fermentation Sciences David H. Koch School of Engineering Practice, MIT Significant contributions by Lindsay Arnold 24

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