Operating Instruction & Manual Lachesis Controller s

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Operating Instruction & Manual Lachesis Controller s Document date: 2017-01-20 Version 1.

1 Operating Instruction & Manual Lachesis Controller s Document date: Version 1.5 make technical changes to the equipment or changes to this document without any prior notice. - P age 1 28

2 Contents 1. Pre-face Product purpose Device Overview Lachesis internal design Requirement Specification Technical setup You can program these parameters Details to how measurements is done Support for CFF from SpectrumLabs Start-up Fault information Communication Instruction Set up Lachesis, pressure, vacuum, sensor, pump and power Turn on device Set time Pump selection Fluid flow, Cleaning/Harvest and Straws & ID Close configuration Turn on and check pressure and vacuum Manually apply pressure and vacuum Run and stop PID routines Operation Manual Operation principles Fluid diagram Safety precautions Documentation make technical changes to the equipment or changes to this document without any prior notice. - P age 2 28

3 1. Pre-face Lachesis is a series of control stations designed for operation of Clio diaphragm Single-Use-Pump (SUP) integrated with the CellMembra P-SUB and a Cross-Flow-Filter (CFF) for cell retention. 1.1 Product purpose CellMembra products is a range of Perfusion-Single-Use-Bioreactor s (P-SUB s) for high cell density cultivation operating in perfusion mode. Such as cell retention system for upstream Continuous Processing, cultivation of mammalian cell lines as supplied by The P-SUB integrates both Clio Single-Use-Pump (SUP) and stacked hollow fibre module / Cross-Flow-Filter (CFF) - all in one package. The process is also named PTF Pulsating-Tangential-Flow. Driving the diaphragm Clio SUP requires drive gas both below and above atmospheric pressure. The drive gas are constantly adjusted via a set of proportional valves inside Lachesis. Control is based on input from sensors (distance and pressure) given to the build-in CPU. Stroke and number of strokes determine the pumped broth volume and velocity of the broth passing through the CFF. 2. Device Overview The Lachesis units is a standard product manufactured by housed in a Hephaestus U2 cabinet manufactured from AISI 304 stainless steel. For single channel Lachesis-1 front panel is equipped with: 1. 5 touch sensitive TFT display 2. Main power breaker 3. Buccaneer 24 VDC power input 4. USB socket for Wi-Fi access and charging 5. RJ45 socket for Local-Are-Network (LAN) connection 6. SUP laser sensor input with M12, RKF5, 5P socket 7. Supply pressure 6 mm one-touch connection 8. Supply vacuum 6 mm on-touch connection 9. SUP drive via 6 mm hose and on-touch connection make technical changes to the equipment or changes to this document without any prior notice. - P age 3 28

For dual channel Lachesis-2 front panel is equipped with: 1. 5 touch sensitive TFT display 2. Main power breaker 3. Buccaneer 24 VDC power input 4. USB socket for Wi-Fi access and charging 5.

4 For dual channel Lachesis-2 front panel is equipped with: 1. 5 touch sensitive TFT display 2. Main power breaker 3. Buccaneer 24 VDC power input 4. USB socket for Wi-Fi access and charging 5. RJ45 socket for Local-Are-Network (LAN) connection 6L. Left channel laser sensor input with M12, RKF5, 5P socket 6R. Right channel laser sensor input with M12, RKF5, 5P socket 7. Supply pressure 6 mm one-touch connection 8. Supply vacuum 6 mm on-touch connection 9L. Left channel SUP drive via 6 mm hose and on-touch connection 9R. Right channel SUP drive via 6 mm hose and on-touch connection There are two fan openings on the rear panel. Avoid to push the cabinet close to a wall prohibiting ventilation and internal cooling. 2.1 Lachesis internal design Internal sensors and actuators: Pressure sensor for supply, >1 - <4 bar atmospheric (will tolerate up to 8 bar supply) Vacuum sensor for supply, <0,1 bar absolute (will tolerate up to 8 bar supply) First channel drive gas pressure / vacuum sensor outlet to SUP, ± 1 bar Second channel drive gas pressure / vacuum sensor outlet to SUP, ± 1 bar Proportional valve for positive drive gas pressure regulation Proportional valve for negative drive gas pressure regulation Fan for cabinet temperature control make technical changes to the equipment or changes to this document without any prior notice. - P age 4 28

5 External sensors needed: For each channel a re-usable tri-angular Leuze laser sensor mounts on top of the re-usable bracket on the SUP for on-line measurement of the actual position of the free-floating silicone membrane / diaphragm inside Clio SUP. 2.2 Requirement The unit must be properly installed according to fluid diagram and general recommendations. 2.3 Specification When Lachesis is connected to: a suitable supply of pressurised air with sufficient capacity a suitable supply of vacuum with sufficient capacity Leuze laser sensor(s) Clio Single-Use-Pump(s) then the spec is: Stroke frequency, per minute 0,1 20 depending on SUP size Stroke duration, seconds, seconds 3-60 Stroke volume Clio ml Stroke volume Clio ml Stroke volume Clio ml Max pumped volume Clio-30 Max pumped volume Clio-100 Max pumped volume Clio-150 ~ 0,5 litre/min ~ 1 litre/min 1,5 litre/min ~ 1,5 litre/min CFF / straw internal velocity, m/s 0,1 >10 GUI 5 touch sensitive TFT display Computer power 900 MHz quad-core ARM Cortex-A7 CPU running Linux with Code-Sys PLC software USB socket Wi-Fi like NetGear N300 Mini Adaptor even charging 5 VDC 1 amp RJ45 socket IP/TCP via LAN (ModBus protecol) Power supply 24 VDC, >30 watt via Buccaneer socket Build in fan, noise level, dba <65 Duty cycle 100% Orientation horizontal Operating conditions 10 C to 40 C, <80% relative humidity non condensing Life time, estimated, hours <50,000 10,000 MTBM (mean time between maintenance) clock battery must be changes, hours Cabinet size and material Hephaestus U2 AISI304 Weight, kilo 3,5 make technical changes to the equipment or changes to this document without any prior notice. - P age 5 28

6 3. Technical setup Figure 1: Graphical-User-Interphase for Lachesis controller as delivered by build in webserver. 3.1 You can program these parameters Press in each section of the Light green area and you will through up a keyboard. Cleaning / Harvest How many harvest cycles between if any cleaning cycle. You can obtain double or triple velocity in cleaning cycle. You must inset 0 and accept in order to change any new setting. SP flow The re-circulated volume in ml per minute for the PTF process ml Number of straws Most often 12 straws in the CFF systems. Only 1 mm inner diameter. Select 0-99 straws. Clock config For the time being manually inserted clock time. Future will be process duration, the planned duration time for the process based on internal clock. make technical changes to the equipment or changes to this document without any prior notice. - P age 6 28

Figure 2: Lachesis programming and input window Run Start up the process based on latest program. Stop Terminate the process based on latest program.

7 Figure 2: Lachesis programming and input window Run Start up the process based on latest program. Stop Terminate the process based on latest program. Vacuumize Button for manual and upwards and slow SUP piston movement. Use for system priming. Pressurise Button for manual and downwards and slow SUP piston movement. Use for system priming. 3.2 Details to how measurements is done Stroke frequency, maximum per minute: 1-10 How many strokes over time is calculated by Lachesis. Could be one stroke per minute or as much as 10 per minute. Limited by general friction, straw deposits, drive gas pressure / capacity and broth viscosity. Stroke duration, seconds: 8-60 Piston travel, reciprocate between two selected positions. Max piston travel distance is 180 mm. Pumped volume, liter per minute, LPM: 0-2 or 0-5 SUP displacement and stroke frequency determines the total volume over time. The SUP with reciprocating piston will pump double the programmed volume as the pump stroke is 50% of the time. The display show programmed curve and the actual curves. If the programmed pumped volume is too ambitions the display will show. Pumped volume per stroke, ml: ~160 or 350 SUP displacement, volume to be pumped is fully dynamic and measured on-line. CFF velocity, m/s: Velocity of piston gives the corresponding velocity of broth passing the hollow fiber tubes in the CFF. Depending on the CFF module the pump gives different CFF velocity. End-user must program in the number of straws. Calculation is based on 1 mm straws. make technical changes to the equipment or changes to this document without any prior notice. - P age 7 28

Figure 3: Lachesis Data acquisition window. The data show should be self-explanatory. Its good to know that Real-time flow is approximately the double of Average-flow.

Data from drive gas is shown online so it the actual pressure above the piston. Jumping between vacuum and pressure.

8 Figure 3: Lachesis Data acquisition window. The data show should be self-explanatory. Its good to know that Real-time flow is approximately the double of Average-flow. This because the SUP is a reciprocating pump and fluid is moved only at the down wards stroke. Total volume moved can only be from process start by pressing Run button. Data from drive gas is shown online so it the actual pressure above the piston. Jumping between vacuum and pressure. Alarms for: Alarm area will show if the supply hose for pressure and vacuum is not mounted correctly. Pump pressure range, Bar: 0.5 to 0.9 Supply pressure range, Bar: 0.5 to 0.9 Vacuum pressure range, Bar: 0.5 to 1.0 In general if no pressure or vacuum is available or drops out then an alarm and process stops. 3.3 Support for CFF from SpectrumLabs Cronus-PCS Lachesis supports different dimensions of hollow filter modules. Both available manufactured from modified Polyethersulfone (mpes) typically with 750 kd 0,2 µm pore and Polyethersulfone (PES) with 0.2 µm pore. The chart show data and physical sizes. Table 1: Hollow filter modules. make technical changes to the equipment or changes to this document without any prior notice. - P age 8 28

9 4. Start-up The scope of the product is to control the function of a SUP connected to a P-SUB. The SUP consist of a tube, cylinder with a reciprocating piston. The SUP being connected on its inlet to a SUB and on the outlet to a CFF. This in order for a controlled volume of the broth to be forced through the CFF. After unpacking and disposing the shipping material properly the Lachesis unit is ready for installation. The unit is pre-programmed with a general procedure according to the Operation Manual section. Important procedure to follow: 1. Connect a pressurised drive gas supply of max 1 Bar (such as Sarpedon / Alagonia) with 6 mm rigid hose. 2. Connect a vacuum source (such as Sarpedon / Alagonia) with 6 mm rigid hose. 3. Connect the rigid 6 mm drive gas hose from the SUP sterile filter to Lachesis. 4. Mount the level sensor onto the SUP top on CellMembra. 5. Connect 24 VDC from one of the Eos family of power supplies. 6. Start-up Lachesis and watch the unit get alive 7. Spend some minutes and use the two manual buttons to remove air and fill SUP and CFF with liquid. Insure the broth is returned to the SUB 8. Program according to you SOP and planning 9. The Lachesis is ready for work Requirement The system must be properly installed and connected in accordance with the specifications and previous information. Operator must also have gained familiarity with the Safety Instructions to be found separately on Fault information The build-in display of Lachesis will inform about possible faults whenever detected. If no drive gas pressure or vacuum source is detected If no movement is detected by level sensor 5. Communication Lachesis contain a webserver displaying online information on the build-in display. Each Lachesis product has an IP address shown at the Manufacturer s Identification Label under the product. If you hook up via the Wi-Fi connection to a touch sensitive PC, smartphone, or PAD you should see the GUI from the actual Lachesis unit. 5.1 Instruction This section has subsections that explain the steps very detailed how to set up Lachesis. 1) Make sure every power supply, hose, pressure regulator, pressure gauge, laser sensor and USB Wi-Fi component is connected to the right places, it is recommended to be sure about the device overview. 2) When the setup is ready, then turn on the Lachesis. 3) Put in the Date and time. 4) Select pump. 5) Put in the data for cleaning/harvest relation, SP flow and velocity calculation. 6) Close configuration. make technical changes to the equipment or changes to this document without any prior notice. - P age 9 28

7) Turn on pressure and vacuum and check the values around 1 bar and -1 bar. 8) Manually apply pressure and vacuum to secure that air in the pump is replaced with fluid. 9) Run the process. 5.1.1 Set up Lachesis, pressure, vacuum, sensor, pump and power.

has to be connected for both Lachesis-1 and Lachesis-2, below there a picture of how it could look like, these setups are with our own Sarpedon reservoir and Alagonia pressure/vacuum 1 bar pumps.

10 7) Turn on pressure and vacuum and check the values around 1 bar and -1 bar. 8) Manually apply pressure and vacuum to secure that air in the pump is replaced with fluid. 9) Run the process Set up Lachesis, pressure, vacuum, sensor, pump and power. Before the Lachesis and the pressure/vacuum is turned on it is important to check that everything is in the right places, under the section Device overview there is a picture showed where everything has to be connected for both Lachesis-1 and Lachesis-2, below there a picture of how it could look like, these setups are with our own Sarpedon reservoir and Alagonia pressure/vacuum 1 bar pumps. Figure 4: Lachesis-1 set up. Figure 5: Lachesis 2 set up. make technical changes to the equipment or changes to this document without any prior notice. - P age 10 28

11 5.1.2 Turn on device Secure that the power cable is plugged in and the switch is turned on, and then press the main power breaker (nr.2 on the device overview of Lachesis) so it will turn on a white light where the blue circle is on the figure. Lachesis is on: Set time First thing to do is to set the date and time which is done by pressing these light green areas. Figure 6: Lachesis set date and time bar. make technical changes to the equipment or changes to this document without any prior notice. - P age 11 28

The way it is done is by one pressing a field at a time, which allows access to insert the desired value. For example if you press Y which is the name for years this window will pop up.

3 - Clear that erases all numbers. 4 - ESC is the escape button that will allow you to close down the pop up window without any changes.

12 The way it is done is by one pressing a field at a time, which allows access to insert the desired value. For example if you press Y which is the name for years this window will pop up. Figure 7: Pop up window. Figure 8: Pop op window description. The pop up window contains: 1 - Numbers that is used to put in the desired value. 2 - Back that erases number rightmost. 3 - Clear that erases all numbers. 4 - ESC is the escape button that will allow you to close down the pop up window without any changes. 5 - OK will set the desired value into the system and close the pop up window if the value is within the range of minimum and maximum values. When the years, months, days, hours, minutes and seconds are keyed in, it should look like this. Figure 9: Lachesis date and time bar with new values. If all values are as desired then press the Set Time field to accept the changes in the system. Figure 10: Lachesis set time button make technical changes to the equipment or changes to this document without any prior notice. - P age 12 28

13 This will set the Actual time in the system to start counting from the Wanted time and when done right it will look like the picture below. Figure 11: Lachesis with date and time set. make technical changes to the equipment or changes to this document without any prior notice. - P age 13 28

14 5.1.4 Pump selection to select the pump size you have to look under Pump selection and press the field that say Disabled. Figure 12: Lachesis disabled button Now a dropdown menu will appear and show a selection of pump size to select. Figure 13: Lachesis with pump selection dropdaown menu open. When the size of pump are decided simply press at the grey field of where it appears and it will if we select Clio100 look like the picture below. Figure 14: Lachesis Clio100 pump selected. make technical changes to the equipment or changes to this document without any prior notice. - P age 14 28

5.1.5 Fluid flow, Cleaning/Harvest and Straws & ID Now we concentrate around this area of the screen where fluid flow, Cleaning/Harvest and Straws & ID value is put into the system.

15 5.1.5 Fluid flow, Cleaning/Harvest and Straws & ID Now we concentrate around this area of the screen where fluid flow, Cleaning/Harvest and Straws & ID value is put into the system. Figure 15: Lachesis, cleaning/harvest relation, SP flow and Velocety calculation fields. For now we look how to select the millilitre flow per minute, press the light green field in the red area in the picture below. Figure 16: Lachesis SP flow selection. After pressing the light green field this pop up window below will appear like when setting the time. Figure 17: Lachesis SP flow pop up window. make technical changes to the equipment or changes to this document without any prior notice. - P age 15 28

The cleaning/harvest relation you set the right value like 1:50. Before After Figure 19: Cleaning/Harvest before set.

16 This will again show the minimum and maximum mmmm that can run through the pump, after inserting mmmmmm the value press OK to confirm. After pressing OK the value will appear in the red square. Figure 18: Lachesis with SP flow set. The cleaning/harvest relation you set the right value like 1:50. Before After Figure 19: Cleaning/Harvest before set. Figure 20: Cleaning/Harvest after set. make technical changes to the equipment or changes to this document without any prior notice. - P age 16 28

Figure 21: Lachesis velocity calculation pop up window. Before After Figure 22: Velocity calculation before set.

17 The Velocity calculation is a little more difficult because you have 2 variables that can be changed, the left one is like we know it, but for the mm value you need to know that the dot is placed in the bottom right corner of the pop up window. Figure 21: Lachesis velocity calculation pop up window. Before After Figure 22: Velocity calculation before set. Figure 23: Velocity calculation after set. make technical changes to the equipment or changes to this document without any prior notice. - P age 17 28

18 5.1.6 Close configuration Now that we got every value into the system you are ready to watch the system in full scale therefore you close down right side of the screen to show how the system is running. You close it down by pressing the close field under the configuration. Figure 24: Lachesis, configuration open. And now it will show it like this: Figure 25: Lachesis, configuration closed. make technical changes to the equipment or changes to this document without any prior notice. - P age 18 28

19 And if the laser sensor is connected it will make a straight line because the membrane is not moving. Figure 26: Lachesis with laser sensor connected. make technical changes to the equipment or changes to this document without any prior notice. - P age 19 28

20 5.1.7 Turn on and check pressure and vacuum Turn on the power of the pressure and vacuum supplies. The way to check that the pressure and vacuum values are correct is to look of the Sup. Pressure field and Vac. Pressure field values in the bottom left corner, the Pump Pressure field is indicating how much pressure or vacuum the pump is working with. Figure 27: Lachesis with no added pressure and vacuum. Figure 28: Lachesis with the correct Pressure and Vacuum added. make technical changes to the equipment or changes to this document without any prior notice. - P age 20 28

5.1.8 Manually apply pressure and vacuum Before running a program it is suggested that you manually pump the air out and the fluid in so that the controller find the ideal line faster.

21 5.1.8 Manually apply pressure and vacuum Before running a program it is suggested that you manually pump the air out and the fluid in so that the controller find the ideal line faster. So the way the pressure and vacuum is manually controlled is by pressing these two light green fields that is shown on the picture below. Figure 29: Lachesis witch manual pump control focus. First remove the air by pressing the pressurise field and it will make your graph go up because the membrane is going away from the sensor like it is show on the picture below. Figure 30: Lachesis where pressure is added to Clio100. make technical changes to the equipment or changes to this document without any prior notice. - P age 21 28

22 Then pump some fluid in by pressing the vacuumize field, it will make your graph go down because the membrane is getting closer to the sensor like it is show on the picture below. Figure 31: Lachesis where vacuum is added to Clio100. Repeat it 3-5 time or until the pump is filled only with fluid. make technical changes to the equipment or changes to this document without any prior notice. - P age 22 28

23 5.1.9 Run and stop Next step is to make the system run automatically, that is done by the automatic pump control where both run and stop field are shown. Figure 32: Lachesis where automatic pump control is in focus. If every parameter is in the system, it is ready to run, that is done by pressing the run field and an ideal line will appear, the picture below show how it look like. It is important to notice that there is no pressure or vacuum on the picture, which are showed in the left bottom. Figure 33: Lachesis that run without pressure or vacuum added. make technical changes to the equipment or changes to this document without any prior notice. - P age 23 28

24 If the pressure and vacuum is on it will follow the ideal line and look like the picture below, where it is showed that the dark line (laser sensor) is following the ideal line in the program: Figure 34: Lachesis that run with pressure and vacuum added. And it is easy to see how the controller at any time will try to aim at the ideal line, so the curve is nice and what is wanted of the user. When the Lachesis is done with is work or have to have other parameters the stop field next to the run field is pressed. make technical changes to the equipment or changes to this document without any prior notice. - P age 24 28

25 5.2 PID routines Many hours has been invested in setting up the PID routines and the intelligence for auto-tuning, which means that Lachesis will align with the inserted liquid flow. It s of importance that you use the two manual buttons Vacuumize and Pressurize to remove all air and fill up with liquid. Watch the full piston movement and see liquid is r-circulated to the SUB. If all air is not removed and liquid not fully filled it will take longer time for the auto-tuning to figure out what s going on! 6. Operation Manual At activation of power button then Lachesis (or restoration of voltage after a power outage) starts up and the display will show: The operating system software (Linux) is loaded The system configuration is loaded. The operating software is loaded and actual information shown on display Any user-defined parameters from a previous process are stored in a battery-buffered memory and can be used for the next process. Lachesis is pre-programmed from the manufacturer with simple routines. If there is no faults are found by the software is ready and ready to operate The GUI is divided into 4 groups: Light green area with 8 buttons Dark green area with information Large graphical chart Small elongated graphical chart 1. Frequency or Ratio between harvest and cleaning cycles 1: Set-Point of flow, ml/min: choose max 3. Number of straws in the CFF for velocity calculation: Clock config insert the actual time 5. RUN start-up the process 6. STOP terminate the process 7. Vacuumize: manual operation by pushing the button (use before start-up) 8. Pressurize: manual operation by pushing the button (use before start-up) make technical changes to the equipment or changes to this document without any prior notice. - P age 25 28

26 Illustration of the webserver offered GUI shown on the build-in 5 display. 6.1 Operation principles Assuming a correct assembly of all systems, connections, etc. according to Fluid Diagram. The SUP is arranged between SUB and CFF. On each side a passive valve insure broth flow in one direction only. From SUB through SUP to and through CFF back to SUB. The free flowing piston inside the SUP cylinder separates the broth from the drive gas. The laser sensor read at any time the position of the piston. Velocity of piston is controlled by volume of drive gas and travel distance. Drive vacuum insure piston is dragged towards the SUP top sucking broth from the SUB. Broth volume and hereby filling displacement is measure by laser level sensor and controlled by Lachesis. Drive pressure force piston towards SUP bottom and force broth to and through CFF. Pumped broth volume pass through CFF to the retentate side of CFF and is returned to SUB. make technical changes to the equipment or changes to this document without any prior notice. - P age 26 28

27 6.2 Fluid diagram The following fluid diagram is an example Illustration - Fluid diagram as setup and method of operation. 1. Stroke frequency, maximum per minute: 10 How many strokes over time, determined by Lachesis. Could be one stroke per minute or as much as 10 per minute. 2. Stroke duration, seconds: 1-60 Piston travel between two selected positions over time and time in between strokes determines the velocity. Max travel distance is 180 mm. 4. and 6. Pumped volume, liter per minute, LPM: SUP displacement and stroke frequency determines the total volume over time 3. and 5. Pumped volume per stroke, ml: SUP displacement, volume to be pumped is fully dynamic and measured on-line. 7. CFF retentate velocity, m/s: Velocity of piston gives the corresponding velocity of broth passing the hollow fiber tubes in the CFF. Depending on the CFF module the piston velocity gives different CFF velocity. Calculation is required. Picture????? Illustration of the principles two different cycles, one for harvest and one with 5 times the volume and 3 times the velocity. make technical changes to the equipment or changes to this document without any prior notice. - P age 27 28

28 7. Safety precautions Various component require individual attention. Operator must also have gained familiarity with the Safety Instructions to be found separately on Documentation Lachesis functionality must be checked on a regular basis and data of such testing kept recorded. END make technical changes to the equipment or changes to this document without any prior notice. - P age 28 28

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