Multi-column Continuous Capture Chromatography New data confirming significant savings and de-bottlenecking opportunities Dr.-Ing. Kathleen Mihlbachler, LEWA Bioprocess Repligen Seminar: Advances in Bioprocessing, Princeton, NJ, 20 June 2017
Agenda Who we are, what we do Drivers to multi-column capture chromatography Streamline twin-column system Verification at process scale during customer trials LEWA-Nikkiso America Inc. 2
LEWA Bioprocess Group is part of the global family of companies under the $1.3B US Nikkiso Corporation (1953) Business sites in >80 countries, >6,000 employees LEWA Bioprocess Group Engineering Fabrication LEWA-Nikkiso America, Inc. Sales Engineering Fabrication LEWA GmbH Headquarters Industrial Division Sales Engineering Pump production Fabrication Nikkiso Co., Ltd. Group Headquarters
Who we are, what we do LEWA Bioprocess Technologies Group A group of ~ 60+ professionals dedicated to GMP scale bioprocess systems design and manufacture. Part of the ~ 6,000 employees of Nikkiso Corp. a diversified company with $1.3B in revenue. Location Date of Establishment Ownership About LEWA GMP systems engineering, manufacturing & automation; Chromatography Buffer In-line Dilution TFF Downstream process skids Markets: Biopharmaceutical Pharmaceutical OEM (bio/pharma) Devens, MA USA 2000 Acquired in 2014, we are now a group within LEWA-Nikkiso America, Inc. The LEWA group ($280M US) is a manufacturer of metering pump headquartered in Leonberg, DE. LEWA was acquired by Nikkiso in 2008. Both Nikkiso & LEWA founded ~65 years ago Video Link Here 4 7/10/2017 7/10/2017 4 LEWA Process Technologies, Inc.
Agenda Who we are, what we do Drivers to multi-column capture chromatography Streamline twin-column system Verification at process scale during customer trials LEWA-Nikkiso America Inc. 5
A paradigm shift to continuous bio-production Drivers Third party payer cost pressure driving need for higher productivity Improved ratio up/down time Biosimilar/biobetter demand to be more cost effective Space constraints: closed processing without hold-up tanks at smaller scale Flexibility when scaling-up to scale out FDA encouraging implementation of continuous processes: Increase quality, reduced production risks Make drugs more available to more populations 6
Why continuous chromatography for capture? Harvest Feed Bleed Eluent Upstream Perfusion Culture Continuous capture Drivers to transfer capture to continuous operation: Breaking the bottleneck in DSP due to higher titers and continuous upstream Volume reduction of harvest - Space constraints Scale down of conventional LPLC systems Elimination of large buffer and hold-up tanks Scale down of columns (required for traditional capture up to 250 L) Under utilization of Protein A binding capacity (30-50%) 7
Transition from batch to continuous capture Single column batch chromatography Load 30-50% unused resin capacity Two-column continuous batch chromatography Interconnected operation Load Parallel operation Recovery Product elution Load Higher resin utilization > 90% OpEx reduction Higher feed and process flow rates higher throughput Reduced buffer consumption OpEx reduction 8
Agenda Who we are, what we do Drivers to multi-column capture chromatography Streamline twin-column system Verification at process scale during customer trials LEWA-Nikkiso America Inc. 9
Simplicity 8 column process CUBE from ChromaCon LEWA EcoPrime Twin ~240 Valves ~40 Valves The 2-column approach reduces risk Accelerates validation Simplifies maintenance Lessens costly downtime Minimizes hold-up & operating expenses Streamlines implementation; easy-tounderstand and use Similar productivity as systems with 3 or more columns, in a simple 2-column design. and 10
Process automation example HMI Main screen (EcoPrime Twin System) Process flow chart with sensors to monitor and control process. Trends, Sequences and Recipes, Reporting and Alarms User trained on day 1 and up and running on day 2 Connected I2: Column 2 in first position 11
Continuous Capture using the 2-column system CaptureSMB Process Patent by ChromaCon www.chromacon.com 12
Recipe driven process configuration (EcoPrime Twin 100) Access control Second person Verification, different recipe stages: Development, Production and Archived 13
Agenda Who we are, what we do Drivers to multi-column capture chromatography Streamline twin-column system Verification at process scale during customer trials Skid performance Customer A 1 g/l mab Customer B 5 g/l mab LEWA-Nikkiso America Inc. 14
Extended run time performance verification Long-term study over 210 loads (13 days) (EcoPrime Twin 100) 350 3.0 300 2.5 Feed pump (ml/min) 250 200 150 100 Monitoring feed flow rate and pump discharge pressure (Shown - last 24 hours only) 2.0 1.5 1.0 0.5 discharge (bar) 50 0.0 0 Consistent and robust performance 15
Reproducibility performance verification (EcoPrime Twin 100) Shown: monitoring conductivity at the outlet of both columns Consistent & reproducible performance 16
Simple transfer from batch to continuous capture Customer A - process development run comparison with 1g/L titer mab Batch Capture Total CV 7.85 L Continuous Capture Total CV 1.6 L CV Linear [cm/h] Column Equilibration 3 250 Load 40 150 Wash 1 2 150 Wash 2 2 250 Wash 3 3 250 Elution 3 250 post wash 1 3 250 post wash 2* 2 250 Regen* 3 250 Process time: 9 h CV Load Start-Up 15 Load Connected 27 Load Parallel 22 Wash 1 connected 1 Wash 1 Parallel 1 Wash 2 2 Wash 3 3 Elution 3 Post-Wash1 2 Post Wash 2 1 Regeneration 3 Re-Equilibration 5 Cycle time: 4 h at linear velocity of 250 cm/h Essentially same process steps simple process transfer Higher load volume 40 vs 49 CVs higher resin utilization & reduce buffer consumption Shorter residence times faster loading & shorter processing time 17
Continuous capture reproducibility Customer A 1g/L titer run results at 40 g/l load 7 peaks overlaid Column 1 At steady state, variability in peak area < 0.25 %. Steady state can be reached with first elution. Column 2 Peak Area column1 column2 1 0.153 0.192 2 0.170 0.194 3 0.170 0.193 4 0.170 0.193 5 0.170 0.194 6 0.170 0.194 7 0.170 0.194 Average 0.170 0.193 error % 0.1764 0.2169 EcoPrime Twin 18
Optimized continuous capture reproducibility Customer A 1g/L titer mab: 50 g/l load 5 peaks overlaid EcoPrime Twin Column 1 Peak Area Column 1 Column 2 1 0.250 0.294 2 0.263 0.297 3 0.265 0.297 4 0.265 0.297 5 0.265 0.297 Average 0.264 0.297 error %* 0.639 0.025 At steady state, variability of peak area is <1 %. Steady state can be reached with first elution. Column 2 19
Comparison batch to continuous capture Customer A - process development run: 1g/L titer mab Triple productivity or 67 % less resin and ~ 30% buffer reduction 20
Scale-up estimate Customer A 1 g/l titer estimate for 100-kg process scale-up Savings up to $900K in resin or 22 days in processing time. And 15,000 L of buffer. 21
Seamless transfer from batch to continuous capture Customer B pilot plant process comparison; 5 g/l titer mab Batch capture Load 50 g/l resin Total CV 17.3 L Continuous capture Load 80 g/l resin Total CV 1.6 L CV linear [cm/h] Pre Sanitization 3 300 Equilibration 3 300 Load 10 150 Wash 1 2 300 Wash 2 5 300 Wash 3 3 300 Elution 5 300 CIP 3 300 Neutralization 1 3 300 Sanitization 3 300 Neutralization 2 3 300 Storage 3 300 Process time: 4.5 h CV linear [cm/h] Pre Sanitization 3 400 Equilibration 3 400 Load 4.6 100 Load interconnected 11.4 150 Wash 1 2 400 Wash 2 5 400 Wash 3 3 400 Elution 3.5 400 CIP 3 400 Re-equilibration 3 400 Sanitization 3 400 Neutralization 2 3 400 Storage 3 400 Cycle time: 2.5 h Same process steps simple process transfer Higher load 10 vs 16 CVs higher resin utilization & reduce buffer consumption Faster linear velocity shorter processing time 22
Reproducibility and scalability Customer B 5g/L titer mab: 80 g/l load EcoPrime Twin run with 14-h process time peak # Column 1 Column 2 peak area peak area 1 0.239 0.276 2 0.255 0.283 3 0.259 0.285 4 0.260 0.286 average 0.2583 0.2849 error %* 1.1 0.7 Similar chromatograms when scaling up and down Contichrom Cube run with 12-h process time Steady state can be reached with first elution. Variability of peak area is ~1 % at steady state 23
Comparison batch to continuous capture Customer B - process development run: 5 g/l titer mab Productivity - Customer B Buffer Consumption - Customer B 25 20.6 22.2 0.8 0.73 Productivity [g/lresin/h] 20 15 10 5 8.4 Buffer Consumption [L/g] 0.7 0.6 0.5 0.4 0.3 0.2 0.1 0.36 0.34 0 Batch EcoPrime Twin Contichrom Cube 0 Batch EcoPrime Twin Contichrom Cube > 2X productivity or 50 % less resin and ~ 50 % buffer reduction 24
Scale-up estimate Customer B 5 g/l titer estimate for 100-kg process scale-up Savings up to $320K in resin or 6.1 days in processing times. Plus 37,000L of buffer. 25
Customer experience on mab shows Twin column capture delivers: Significant productivity gains 2-3 times Substantial cost or time savings Better Protein A utilization Faster loading Shorter processing time Reduced buffer consumption..30-50% Seamless scale up Same process steps Proven reproducibility Simple 2-column approach and system Accelerate implementation Minimize risk LEWA Nikkiso America Inc. 28
For more information: www.lewapt.com America s Europe Asia LEWA-Nikkiso America, Inc. 132 Hopping Brook Road MA 01746 Holliston Tel: +1 508 429-7403 Fax: +1 508 429-8615 sales@lewa-inc.com Headquarters LEWA GmbH Ulmer Str.10 71229 Leonberg GERMANY Tel: +49 7152 14-0 Fax: +49 7152 14-1303 lewa@lewa.de NIKKISO CO. LTD. Yebisu Garden Place Tower 22nd Floor 20-3, Ebisu 4-Chome, Shibuya-ku 150-6022 Tokyo Tel: +81 3 3443-3711 Fax: +81 3 3473-4963 LEWA, Creating Fluid Solutions, the LEWA logo, LEWA Intellidrive and LEWA EcoPrime are trademarks of LEWA GmbH. ChromaCon, Contichrom and CaptureSMB are registered marks of ChromaCon AG. All goods and services are sold subject to the terms and conditions of sale of the company within LEWA which supplies them. A copy of these terms and conditions is available on request. Contact your local LEWA representative for the most current information. 2017 LEWA Nikkiso America, Inc. All rights reserved. LEWA Bioprocess Technologies, Inc. 8 Charlestown Road Devens, MA 01434 USA Tel: +978 487 1100 sales@lewapt.com
ChromaCon / LEWA Partnership LEWA is global GMP scale up partner for ChromaCon twin column technology. Control logic for operating two columns on a single system. ChromaCon Process know-how Intellectual property Entry-stage equipment CaptureSMB LEWA Scale-up Equipment Process solutions Continuous manufacturing Worldwide Footprint MCSGP Technical, marketing and sales collaboration. Work protected by various patents 30
Huge productivity System/Model Column ID Range (cm) Feed per day (L/day) Product recovered* (kg/day) EP Twin 100 0.004 0.60 L/min EP Twin 250 0.02 2.4 L/min EP Twin 500 0.06 9.0 L/min 2.5 10 23 360 0.11 1.7 5 20 92 1,460 0.44 7.0 8 45 206 7,420 1.0 35.0 *Process dependent 10 cm ID columns More than 300 L feed processed/day More than 1 kg mab purified/day LEWA Bioprocess Group and 31
Twin column LPLC skid layout EcoPrime Twin 100 2 Flow meters Outlet valve block Column inlet valve block Column outlet valve block 1. Symmetric design 2. Minimum hold-up volume 3. Simple piping design ph probe 2 UV detectors 2 conductivity and temperature 2 ecodos pumps Pump A (Buffer) with 6 inlets Pump B (feed) with 4 inlets Single outlet valve Column inlets and outlets and their by-passes 32