PlantWeb Seminar

1. PlantWeb Seminar Bioreactor Applications

2. How PlantWeb & Rosemount Analytical Address Key Customer Issues

3. How PlantWeb & Rosemount Analytical Address Key Customer Issues

4. How PlantWeb & Rosemount Analytical Address Key Customer Issues

5. How PlantWeb & Rosemount Analytical Address Key Customer Issues

6. Fermentation / Cell Culture • Steam Sterilizable sensor selection critical to fermentation processes • pH Sensors • Dissolved Oxygen Sensors • Instrument selection based upon • Ability to withstand cleaning requirements • Diagnostics to determine sensor’s health • 21CFR Part 11 data retention compliant • Quick, easy start ups

7. Products A B C Feedback control: Product inhibits the first enzyme Why is pH Important? • Configuration and Activity of Enzymes and Protein Catalysts Are pH Dependent and Control Growth Kinetics • Enzymes: Special Class of Proteins That Are Biological Catalysts Which Expedite Reactions by Lowering the Amount of Activation Energy Required Enzymes SPEED up Reactions

8. If the pH readings are outside the tolerance levels, rework may be required OR the batch may have to be discarded costing operations $Millions! pH in Fermentation • Guidelines for pH Control • Bacterial: Maximum 6.5 - 7.5 pH • Yeasts: Maximum 4 - 5 pH • Mold: Maximum 5 - 7 pH • The pH for Maximum Growth Rate Typically Increases With Increasing Temperature

9. Bioreactors / Cell Culture pH Solutions • Model 3800 Steam Sterilizable & Autoclavable pH Sensor, rated up to 140°C • Industry standard PG13.5 Threaded Connection • S8 Connector or VP Electrical connection • Multiple Lengths Available • 120 mm, 225 mm, or 325 mm

10. Model 3800 Improvements • Improved Glass Formulation from PERpH-X Development Project withstands steam sterilization cleaning process • Single Ceramic Junction • Improved Reference Electrode minimizes drift

11. Key Differentiators • Improved Performance after Repeated Exposure to SIP • Tested at 140º C • Faster Response to Real pH Changes • Less Drift • Autoclave @ 125º C • Less Drift • SIP @ 140º C • Material Certificates Standard • Includes Actual readings, not % of Theorhetical

12. No Long – Term Sensor Deterioration ±7mV on average after 50 SS cycles! Steam cycles are at 140°C for 30 minutes. Tested in 4 and 10 buffers after each steam cycle.

13. Fast Responding Sensor Response curves after 50 Steam cycles are at 130°C for 30 minutes.

14. Improved Drift Performance pH readings in simulated cell culture after 45 minute autoclave cycle @ 125°C

15. pH readings in 7 buffer @ 37°C after 30 min 140°C steam sterilization cycle Improved Drift Performance

16. Quality Certificate for Each Sensor • Date of certification • Part # & Serial # • Reading in 4 pH, mV • Reading in 7 pH, mV • Slope (pH 4 ; pH 7), % theoretical • Response Time T98% • Membrane resistance, Meg-ohms • Wetted Materials • Inspector signature or mark

17. Determines fermentor/ bioreactor size and configuration Determines feed stock supply rate Effects heat transfer Provides growth profiling Why is Dissolved Oxygen Important? Too much oxygen is not economical or may create side reactions Too little oxygen may stunt the growth

18. Steam Sterilizable Hx Series Dissolved Oxygen Sensors • Proprietary Membrane design reduces maintenance costs • Designed to withstand 50 SIP cycles, or 4x - 6xmore than industry standard sensors • Minimal Flow Sensitivity provides more reliable Dissolved Oxygen Readings • Corrosion resistant design provides longer life • Surface finish < 16µinch Ra • Variable insertion lengths available

19. Why the Hx Series DO is Better • Proprietary membrane: • Membrane will withstand 30-50 sterilization cycles without any maintenance. Most SIP DO probes require a rebuild after 3-8 cycles • Steel-mesh, double membrane withstands high pressures and temperatures while maintaining quick and stable response • Double membrane is less sensitive to contamination • Allows for minimal flow sensitivity for maximum performance • Glass body protects anode and cathode from contamination • Air compartment allows for expansion and contraction of electrolyte to minimize pumping action • Electrolyte is highly alkaline (pH 13.0) • Aids in reducing the electrolyte quickly and efficiently • Allows for short polarization time for fast start-ups Sensor housing Air compartment Replaceable membrane cartridge Silver anode Glass insulation Electrolyte (pH 13.0) Platinum cathode Steel-mesh, double-layer membrane

20. Hx Series - Improved Resistance to Steam Sterilization Cycles Conventional Dissolved Oxygen Sensors

21. Proven Results Application Notes Quantifiable Results Documented Publishable Articles follow

22. Competitor –Fermentor / Cell Culture • Mettler Toledo Ingold • More sensor configurations • Standardize on Variopol • Dissolved Oxygen membranes fail • Dissolved Oxygen cathodes fail • Replaceable cathode design • Private Labeled electronics • Have Profibus • Foundation Fieldbus offering still weak but their new Model 700 looks good

23. New Projects Each fermentor has two pH and 1-2 dissolved oxygen sensors Some will include inductive conductivity for cleaning Foundation Fieldbus MRO pH sensors replaced frequently DO membranes replaced frequently DO Sensors replaced every 3-5 years Sales Potential Sales Strategy:OEM’s have little influence on sensors at larger accounts. End users have final word. Single Use and Contract Manufacturers will rely on OEM’s more on their inputs. Easy to use transmitters with strong digital implementation, sensor’s ability to last through multiple batches, tighter controls, and total cost of ownership are strong sells

24. Probing Questions • How frequently do you change sensors? • How do you determine whether a sensor will operate throughout the campaign? • What happens when a sensor fails during a batch? • How frequently do you standardize sensors?

25. New Family of Hygienic Sensors: Bioreactor Applications