1. Ultrafiltration Presented By: Amar Kher-Bear Jason ‘The Closer’ Ward David ‘Big Game’ Lotimer Derek ‘The Rock’ MacFadden

2. Outline Process Goal Definition Design Goals Setup Unit Sizing Alternative Options Cost Analysis Questions

3. Process Goal Increase insulin concentration coming from the HPLC unit by a factor of 10 before it enters the crystallizer

4. Definition: Ultrafiltration Operates according to principle of diffusion under pressure –Solutes and water are extracted –Retains macromolecules i.e. Insulin Serves two purposes: –Purification –Concenration Differs from diafiltration: diafilters are generally used for simply purification purposes

5. Design Goals High flux High retention rate Hydrolytically stable & Biocompatible Turbulent flow Small footprint Durable Economically viable

6. Operational Setup Batch, Semi-Batch OperationSeries Operation -Largest tank volume and membrane area required. -Conversion per pass is low, but with multiple passes, virtually any concentration can be achieved -Fresh medium continually added to feed tank -Continual re-pressurization required

7. Membrane Options Polyethersulfone (Polymeric) http://www.whatman.com/repository/images/s7/7_57_292_29 0.jpg http://www.sterlitech.com/products/membranes/polyethersulfone/images/Polyet hersulfoneMembrane.jpg http://www.accuton.com/img/products/c_drivers/CM2_elec2 00.jpg Regenerated Cellulose Ceramic -Excellent hydrolytic stability -High flux, high retention -Wide range of operating pH -Durable and thus economically efficient -Expensive -Susceptible to fouling and deformation -Good flux and retention -Comparatively fragile -Narrow operating pH range -Lower flux than polymer membranes

8. Membrane Modules http://www.millipore.com http://www.np.edu.sg/~dept- bio/biochemistry/aab/topics/minitan.gif Perry’s Chemical Engineering Handbook Spiral Wound Cassette Hollow Tube -Cannot produce turbulent flow with the flowrates at which we are operating -Small footprint -Produces turbulent flow for better medium-membrane communication. -Small footprint -Large membrane surface area -High packing density -Susceptible to structural deformation

9. Filter Sizing List of Assumptions: Reference data accurately reflects flux behaviour for Insulin separation with a 1 kDa Polyethersulfone membrane. The rejection coefficient for the Insulin (~6 kDa) retained by a 1 kDa membrane is 1.

10. Design of Ultrafilters centres upon the flux properties of the membrane. Flux is a function of membrane material, protein type, protein concentration, transmembranous pressure Protein Concentration (g/L)

11. Simple Flux Equation: Simple Area Calculation: Membrane Area = 10.2 m 2 #.5m 2 Cassettes = 21 Multi-Step Area Calculation: Membrane Area = 6.5m 2 #.5m 2 Cassettes = 13 SuperPro Calculated Area: Membrane Area = 2.5 m 2 #.5m 2 Cassettes = 5

12. Costing Capital Costs Operating Costs

13. Alternative Options Ion Exchange and Affinity Chromatography - Eluting and buffering chemicals would introduce additional separation units Evaporation –High temperatures denature Insulin –Utilities cost is three times that of Ultrafiltration

14. Engineering Schematic

15. Manufacturers Donaldson – “Filtration Solutions”, Minneapolis, Minnesota Make several different types of filters from compressed air and water and oil separation, to liquid filters Use hollow tube formation of UF Spintek Filtration, INC. - Los Alamitos, California Tubular Ultrafiltration Membranes from SpinTek provide excellent capabilities for filtering and concentrating difficult process and waste streams. Maintenance and replacement savings, along with reduced downtime Millipore - Billerica, MA (but manufacture around the world) Millipore’s Biomax membranes are well suited for applications requiring high flux, low-protein binding, and harsh chemical cleaning and sanitization. For high-recovery purification, the patented Ultracel PLC membranes offer high retention and improved integrity Pall Corporation - East Hills, NY Use a Microza Hollow Fiber Membrane Systems that are designed to integrate with existing infrastructure or as a new, stand-alone infrastructure package Koch Industries INC. - Wilmington, MA Offer both spiral wound and hollow tube UF’s in a wide range of pore sizes

16. References 1. Curtis, Morris, Safford, “Aqueous Protein Solutions Stable To Denaturation”; http://www.freepatentsonline.com/4783441.html; United States Patent 4783441 (2005) http://www.freepatentsonline.com/4783441.html 2. Encon Wastewater Systems Inc., “Competitive Comparison To Ultrafilters”; http://www.evaporator.com/comp_ultra_filters.html; PSI Water Systems Inc., Manchester, NH (2005) http://www.evaporator.com/comp_ultra_filters.html 3. Ferry J.D., Ultrafilter Membranes and Ultrafiltration; Dept. of Chemisty, Stanford University, California; Journal of Chemical Reviews, Vol.18, No.3 (October 1936) 4. Government of Ontario, “Electricity Prices In Ontario”; http://www.oeb.gov.on.ca/html/en/consumerinformation/2005elecrates.htm#3; Ontario Energy Board (2006) http://www.oeb.gov.on.ca/html/en/consumerinformation/2005elecrates.htm#3 5. Henry J.D., Prudich M.E., Eykamp W. et al., Alternative Separation Processes: Ultrafiltration; Perry’s Chemical Engineering Handbook (Section 22-52) 6. Millipore, Pellicon 2 Filters and Holders: Technical Library; http://www.millipore.com/publications.nsf/docs/ds1210en00; Millipore Corporation, Billerica MA (2004) http://www.millipore.com/publications.nsf/docs/ds1210en00 7. Figure “flat plate ultrafiltration system”, Advanced Analytical Biochemistry – Basic Principles; NgeeAnn School of Life Sciences and Chemical Technology, Singapore; http://www.np.edu.sg/~dept- bio/biochemistry/aab/topics/aab_principles.htmhttp://www.np.edu.sg/~dept- bio/biochemistry/aab/topics/aab_principles.htm 8. Millipore Catalogue. http://www.millipore.com; Millipore 2006.http://www.millipore.com 9. “Process and Specialty Liquid Filters”, http://www.donaldson.com/; 2002-2005 Donaldson Company, Inchttp://www.donaldson.com/ 10. “Filtration Systems”, http://www.idswater.com/water/us/spintek_filtration; Spintek Filtration Inc.http://www.idswater.com/water/us/spintek_filtration 11. “Solutions for Biopharmaceutical Applications”, http://www.pall.com/; Pall Corporationhttp://www.pall.com/ 12. “Ultrafiltration – Filtration Overview”, http://www.kochmembrane.com/; Copyright 2004-2005 Koch Membrane Systems, Inc.http://www.kochmembrane.com/

17. Questions?