Technology update: SmartFactory solutions for continuous ...

1 Technology update : SmartFactory solutions for continuous processing Barbara A. Paldus June 12, 2017. What is Perfusion? Definition: Continuously adding nutrient solutions to a bioreactor while removing waste or product from it. The cell culture media is passed through a separation device and the cells are returned to the bioreactor and the cell free media is removed. This removed media is either waste (in some processes) or contains the product. There are several types of separation devices/systems: - Acoustic Separator - Centrifugal Separator - Cell Cube Separator - Hollow Fiber Cell Separator Cell Retention Methods Sample Sample SU.

2 Bioreactor Tangential flow filtration Alternating tangential Floating filter filtration For any bioreactor filtration For rocking bioreactor SU Filter For any bioreactor formats Flexible, many Not single-use Single-use format different sizes and Improves flux over filter Quick and easy setup porosity membrane Limited scale Acoustic Separator System Acoustic Separator (BioSep) uses high frequency sound waves which cause the cells to be suspended inside the sound chamber. Periodically the sound wave is turned off and the cells either fall back into the vessel or are flushed back. The BioSep controller can be electrically connected to the Finesse system by a dry contact relay to coordinate the turning off of the sound and the reversing of the Harvest pump.

3 Vessel size range: 1L to 200L. Finesse Examples: 15L Glass for Insight Israel. Integration requires custom software Centrifuge Separator System Cell culture is removed from the bioreactor via separation pump and delivered to centrifuge inlet; pinch valves are closed. Centrifuge (Centritech Lab III). concentrates the cells over a set time period. Upon completion, the separation pump is turned off, and the pinch valves adjusted so that the concentrated cells are returned to the bioreactor. Pump adds new media based on the system weight when needed Centrifuge Separator System Integrated native DeltaV.

4 Custom control tower Centritech centrifuge with custom control of pinch valves Integration with SUB/glass bioreactors with custom Finesse code used Vessel size range: Labscale to 1000L SUB. Finesse Examples: Shire, Lexington Finesse RDPD with centrifuge CellCube System CellCube (Corning) has a modular cube that contains a series of parallel plates that are joined to create thin laminar flow spaces. Each culture plate has been treated to ensure dense, uniform cell growth. Media is circulated via the oxygenator back into the CellCube. Media is harvested from the oxygenator also. Vessel size range: Labscale Finesse Examples: Merck USA, Boehringher-Ingelheim VetMedica Cell Cube System Single-use centrifuge scalable to 1000L single-use bioreactor Separation mechanism: cell cube vs oxygenator 1.

5 Cells adhere to cell cube walls; cell cube surface area is scalable 2. Oxygenator provides measurement and control of process parameters 3. Recirculation Pump continuously flushes cells with new media Cell Cube System -Example Seeded 100-layer CellCube module cells total 11 L of cells/mL cell suspension harvested cells total collected Average viability of Hollow Fiber Cell Separator (ATF) System ATF Alternating Tangential Flow (Refine Tech/Repligen) has a plastic diaphragm that is inflated / deflated acting as a pump to move the cell culture back and forth through a Hollow Fiber Cell Filter. Cells are retained by the filter and return back into the vessel.

6 Media passes through the filter and to the collection vessel. The ATF. controller is not part of Finesse control system. Pumps & Pressure monitoring are controlled by Finesse. Vessel size range: 1L to 2000L. Finesse Examples: LG, Crucell, Transgene, Boehringer-Ingelheim, Wuxi-Apptec, etc. Hollow Fiber Cell Separator Tangential Flow (Spectrum Labs) uses a disposable magnetic levitating pump to move the cell culture through a Hollow Fiber Cell Filter. Cells are retained by the filter and circulate back into the vessel. Media passes through the filter and to the collection vessel. The Spectrum Labs controller can be integrated to the Finesse control system.

7 Vessel size range: 1L to 2000L. Finesse Examples: Use Spectrum Labs TFF Filter with Finesse controller not using Spectrum Labs controller. Fixed Feed Rate Control Control can be based on a Fixed Feed rate into the vessel Waste SIC-3. Feed Bleed SIC-1. Harvest SIC-2. FIC. Waste WIC or Bioreactor Media Product ATF ATF. Controller Fixed Harvest Rate Control Control can be based on a Fixed Harvest rate from the vessel Waste SIC-3. Feed Bleed SIC-1. Waste or Product Harvest SIC-2. WIC. Bioreactor FIC. Media ATF ATF. Controller Summary of Perfusion at Finesse Finesse is a market leader in perfusion automation and is driving fully integrated upstream-downstream skids Traditional - ATF Centrifuge Custom SmartCart: Universal Filtration STANDARD CONFIG.

8 EXTENDED CONFIG. 2x shelves 2x shelves 6x filter brackets 6x filter brackets 3x Pendotech pressure xmit 3x Pendotech pressure xmit 2x Redlion Freq. xmit for flow 2x Redlion Freq. xmit for flow 3 channel pressure 3 channel pressure 2 channel flow 2 channel flow 1x Temperature (omega Tx) 1x Temperature (omega Tx). 2x mixer connections via Remote 2x mixer connections via Remote VAB. VAB panels panels 1x VFI 1x VFI. 1x Load cell interface 1x Load cell interface 6x pneumatic pinch-type valves 6x pneumatic pinch-type valves 4x Aux Loops (0-10 VDC) for 4x Aux Loops (0-10 VDC) for external external pumps pumps 2x 4-20mA AI's 2x 4-20mA AI's 2x VAC Recepticles (non-ESTOP) 2x VAC Recepticles (non-ESTOP).

9 2x VAC Recepticles (STOP) 2x VAC Recepticles (STOP). Pumps 2x WM520 pumps for pH control 1 x pH/Conductivity 1x rosemount 1056 analyzer pH/Cond OPTIONS OPTIONS. quattroflow 1200 pump quattroflow 1200 pump Max qty = 4 Max qty = 4. Max pressure = 4 bar Max pressure = 4 bar Watson Marlow 720 Pump Watson Marlow 720 Pump Max qty = 4 Max qty = 4. Max pressure = 2 bar Max pressure = 2 bar VAB panels VAB panels Max qty = 2 Max qty = 2. Pressure Transmitters Pressure Transmitters For single use sensors For single use sensors Max qty = 4 (for system total of 7) Max qty = 4 (for system total of 7). Local HMI Local HMI.

10 TruViu Control Panel TruViu Control Panel (1 required per area) (1 required per area). The Aim of Modular Automation Modular approach to production gaining popularity in the Process Industry Improving Efficiency more capable, intuitive skids Increasing Flexibility build-as-you-go and plug-and-play modules Reducing Time to Market COTS solutions with greatly reduced validation cost Synergies between Modular Automation and Single Use Technologies Modular Production will help to master the challenges of the Process Industry Traditional to Modular Automation < 2010 Today Modular design = new approach MES.