Quanta Sep Scale Up Sanitary Automated Chromatography

Quanta. Sep ® Scale Up Sanitary Automated Chromatography Systems

Why Automate your Chromatographic process • • Decrease down time Increase productivity Decrease time to market Consistency in performance Complete documentation GMP and GLP Regulations Economics Sepragen Corporation 2

System Specifications Sepragen Corporation 3

Lab Quantasep Product Line Integrated, bench top, rugged, sanitary- Roche, Amgen, Lonza, SAFC, Hyclone, Invitrogen, Siemens Hardware Feature Set: • Two Pumps (Rotating Piston, positive displacement) • 12 inlets and 10 fractions • 3 columns • Forward flow, reverse flow and bypass Quantasep 100 (100 ml/min max flow) 0 • UV, p. H, temp and Conductivity detection • Sensors for pressure, air, leak and rpm check • Stainless Steel Fluid-handling compartment • Biocompatible materials of construction • Outputs for remote data logging • Fraction collector interface trigger Quantasep 1000 (1000 ml/min max flow) • Emergency pause Sepragen Corporation 4

c. GMP Pilot and Production Quanta. Sep Sanitary, Bio-compatible Systems at Schering Plough, Teva, Baxter, Dyax Quanta. Sep 5000: 5 L/min Quanta. Sep 1000 LX: 2 in one Quanta. Sep® 100 ml/min & 1000 ml/min Quanta. Sep® 5000 Quanta. Sep® 1800 Quanta. Sep 1800: 1800 ml/min Quanta. Sep® 1000 LX Quanta. Sep 10000: 10 L/min Compact foot print; no metals; removal tubing; low system volume. Sepragen Corporation 5

Custom Quantasep Process Chromatography Skids 5 LPM & 30 LPM SKIDS at Human Genome Sciences Custom, Sanitary, 10 -40 Liter per Sepragen Corporation minute

Quantasep Process Chromatography Skids at GSK, Novartis, Neose • • Gradient 2 Pump 10 -40 Liter per minute per pump, 5 -95% Gradient 10 Inlets, 2 Outlets, Filter, Bubble Trap, Column fwd, Rev & Bypass Pre and post column p. H and Cond sensors, Multi wavelength UV Sanitary 316 L SS Piping, 304 SS Moblie Ex Proof Skid Air sensor, Auto air removal and Bubble Trap with control system Leak sensor, Pressure Sensor Filter, Flow through and Bypass CE, BASEEFA (class 1 Div 1, T 4) Certified ATEX explosion proof skid Sepragen Corporation

Dramatic Space savings with Quantasep Quanta. Sep GE Bio-Process 3 mm skid Sepragen Corporation 8

System Selection vs. Column Size Sepragen Corporation 9

Purification of MAb on. Quantasep 100 A simple, load, wash elute and strip Ab isolation using Protein A Column Sepragen Corporation 10

Critical Software features n Intuitive method builder n Dashboard Control n Comprehensive run documentation and report Sepragen Corporation 11

Quantasep Software Feature Set Win XPbased computer control • Easy point & click interface • Auto & manual operation • Method builder and editor • Chromatogram and analysis • Event and alarm log • Easy software calibration • GMP Reports • Part 11 Compliance Sepragen Corporation 12

Software module for Quantasep Systems Capabilities • Real time point& click process control • Data acquisition • Analysis • FDA compliant Reports reports Sepragen Corporation

Manual gradient Sepragen Corporation

Qsep Auto Gradient Run Sepragen Corporation

P&ID with pre-column filter and sensor options Positive displacement pump Pre column filter Filter pressure Inlet Buffers Advanced Gradient System Column Pressure Multimode detector Fraction Collection valves Pre column Multimode detector Active Air Trap Column hookup with forward and reverse flow capability Sepragen Corporation 16

Chromatogram Manipulation And Instant Data Analysis Gradient Instant Data Analysis (buffer consumption etc. during elution Overlays Sepragen Corporation 17

Chromatogram and data analysis Sepragen Corporation 18

Peak Identification and analysis Sepragen Corporation

Intuitive Method Builder n Tabular format - eliminates the hassle of programming n Pick and choose method steps for unprecedented control n Automatic method validation Sepragen Corporation 20

Software Features Easy operation, monitoring& control • Operation from graphical P&ID screen • Manual or Automated Modes • Real-time Chromatogram • Programmable based on time, volume or sensor values • Real time event log • Real time alarms Sepragen Corporation 21

Reports n Validation 4 track alarms, events, notes and any manual changes n Automatic analysis report n Calibration Report 4 Calibration data 4 Time between calibrations n Method and column information Sepragen Corporation 22

Calibration History and Report Sepragen Corporation 23

More Software Features • • • Password controlled security system Chromatogram replay and zoom capabilities Linear or step gradient programmability Generic file formats for easy export Settable alarm conditions Built-in audit tracking capability Sepragen Corporation 24

Product Features and Benefits Sepragen Corporation 25

Differentiating Features §Hardware –Fully Integrated system –Compact foot-print –Real time air ejection –Leak Detect – multimode detector –Pump stall feed-back §Software –Intuitive method builder –Easy P&ID Control –Comprehensive reports –Chromatograms & analysis Sepragen Corporation 26

Precision Rotary piston pumps with stepper motor control • Positive displacement • Stepper motor control • No check valve for good clean-ability • Low pulsation • Small displacement/ rev= even flow Sepragen Corporation 27

Gradient Performance 100: 1 Turn down ratio on pumps Sepragen Corporation 28

Real- time Air Ejection Vs. Passive Bubble Traps Active Air Ejection n Protects against dilution of Product n No Unnecessary Gradient formation n Provides Protection of Column from any amount of air n Can be used to detect end of sample Passive Bubble Trap n Causes Dilution of Product n Unwanted Gradient formation Doesn’t provide complete protection of column against air n Increases Holdup volume Manual air valve Air Outlet Air Ultrasonic air sensor Flow Out Flow In 3 way valve Flow in Flow Out Sepragen Corporation 29

Active Air Trap reduces Elution Volume by 50% With Active Air Trap 0. 25 L elution volume With Standard Bubble Trap 0. 4 L elution volume Sepragen Corporation 30

Cleaning Validation Sanitization Study Process: Inoculate Incubate Wash in Nao. H Water Wash Swab at test points Sepragen Corporation 31

Results of Microbial challenge study on Quantasep 1000 Location S. Aureus E. Coli P. Aerg. C. Albicans A. niger Inlet A 0 0 0 Inlet B 0 0 0 Pump A 0 0 0 Pump B 0 0 0 Manifold A 0 0 0 Manifold B 0 0 0 Pressure 0 0 0 Mixer 0 0 0 Col Outlet 0 0 0 Col. Inlet 0 0 0 Bypass In 0 0 0 Bypass Out 0 0 0 UV flow cell 0 0 0 Cond. sensor 0 0 0 0 0 p. H Ref 0 0 0 Fraction comm 0 0 0 Fraction 1 0 0 0 Air Outlet 0 0 0 Air Inlet 0 0 0 Sepragen Corporation 32

Material of Construction Tubing Silicone, polyethylene and polypropylene, 316 L ss Valves Polypropylene, PTFE diaphragms, 316 L SS Pump Head Ceramic, 316 LL SS Pump Body PVDF, 316 L SS Instrumentation Material glass/quartz/316 L stainless steel Gaskets Silicone, EPDM Sepragen Corporation 33

Multimode Detector Vs. Individual Detectors Multimode Detector • • UV, p. H, Conductivity and Temp. all in one Reduce Lag time between detectors Minimize Hold up / Dead zone More accurate control of Chromatography process Separate Detectors • • Separate UV, p. H, Conductivity and Temp. Too much lag time between detectors. Increases unnecessary Hold up / Dead Zone. Harder to control and validate a process. Flow out Flow in Flow out All Sensors are embedded in one unit Flow in Separate detector flow cells connected by tubing Sepragen Corporation 34

Chromatography Automation Cost-benefit analysis • Overall cost with system – Fixed Cost: System and accessories- manual components – Variable Cost: annual maintenance • Benefits – Direct Saving • Labor Savings in production: Skilled labor (upto $400/hr fully burdened) • Labor Savings in documentation, report writing and admin – Indirect Savings • • • Better time management with 24/7 operation Lesser skilled labor rate can be applied to production Improvement in lot-to-lot consistency or quality Reduced batch failure Accessible documentation for regulatory compliance Sepragen Corporation 35

Cost- Benefit Case Study Incremental investment $258 K Savings/run=25 hrs or$400 x 25= $10 K/run=$30 K/mo Payback= 258/ 30 x 12 =7 mos Sepragen Corporation 36

Competitive Analysis Sepragen Corporation 37

Product Comparison– Qsep 100 vs AKTA Explorer System Positioning Solution/ Price Quanta. Sep 100 is also positioned as small scale production system (simple, compact, Integrated unit design) §Small scale production §Process scale up studies §Integrated design with certain options §Simpler selection §Priced 10 -15% lower than competition §Better features AKTA Explorer is positioned as “all things to all people” tool §Small scale production tool §Process scale up studies §R&D §QC §Price 10 -15% above Qsep 100 §Modular design with “mix and match” of different components §Complex selection criteria Sepragen Corporation 38

Advantage: Quanta. Sep 100 over Akta Explorer Features Qsep 100 Akta Exp Quantasep Advantages Low System Backpressure Yes at max-flow No Akta generates high pressure at 100 ml/min; Qsep is only 50% of the pressure Can Run Pump Dry Yes No Pumps need to be primed or doesn’t flow Pumps don’t siphon Yes No Unintended loss of buffer or sample Auto Air Eject System Yes No Column protected from air intrusion Leak detection Yes No Protection from loss of material Diaphragm valves Yes No Diaphragm valves are more sanitary compared to rotating valves Sanitary design Yes No Sanitary tubing connections; Bug Challenge study Easy to use and program Yes No 2 hrs to learn software as opposed to 2 days Point & Click P&ID operation Yes No Easy to run, monitor and change parameters Real time analysis Yes No Hardware Software Sepragen Corporation Real time superior process control Get buffer volumes; switch conditions; % areas in 39 real time as opposed to post run

Similarities between Quanta. Sep 100 vs Akta Explorer Features Qsep 100 Akta Options Bench top Yes Similar footprint Flow rate 0 - 100 ml/min Similar stated flow range but Akta’s usable flow range is ~40 ml/min Air detector yes Akta uses it to stop flow not remove air dynamically Gradient 0 - 100% Similar Sample injector yes In Quantasep sample loops are standard Detectors UV, Cond, p. H All 3 sensors standard Pumps 2 2 2 Pumps but Akta pumps can only run with primiing Columns 3 1 Quantasep can enable user to do uf/df Manual & Auto Operation Yes Buffer selection; Flow setting; switch conditions based on time, volume, UV, p. H, Cond, Air, Fraction collection Security Yes Controlled access Data Analysis Yes % areas; HETP; buffer usage; switch conditions Reports Yes Method and batch; chromatogram; alarms & events; access Sepragen Corporation 40

Product Positioning – Sepragen Qsep 1000 vs. GE AKTA Pilot System Positioning Solution/ Price Quanta. Sep 1000 is also positioned as small scale production system (simple, compact, integrated unit design) Small scale production Process scale studies Integrated design with certain options §Smaller footprint & more compact §Priced 10 -15%lower than competition §Better features AKTA Pilot is positioned more as a Small scale production Process scale up studies §AKTA “mix and match" multiple components §Complex decision making §Integrated system with few options §Akta priced 15% above Qsep 1000 Sepragen Corporation 41

Advantages Quanta. Sep 1000 vs Akta Pilot Features Qsep 100 Akta Pilot Benefits Buffers 12 8 Greater versatility- enables all 3 columns to be used without stopping run Fractions 10 8 More capability for collecting fractions from multi- column run Low System Backpressure Yes No Operational over 100% range vs. only 60% for Akta Auto Air Eject System Yes No Column protected from air intrusion Leak detection Yes No Protection from loss of material Flow rate/ pump 0 -1000 ml/min 0 - 400 ml/min Greater dynamic range…. . can process large dilute batches faster No Bubble trap Yes No Better chromatography less dilution Point & Click software Yes No Software easy to learn, use and troubleshoot; Akta Software unwieldy to program, train and run overkill for production Real time Analysis Yes No Better process control; lower losses Real time overlays Yes No Increase run- run consistency; Sepragen lower losses and increase yields Hardware Software Corporation 42

Advantages of Quanta. Sep Software vs Competition Software Features Sepragen Amersham Pall Millipore Benefits Point and Click Manual Operation Yes No No No Less training time Online Data Analysis Yes No No No Lower failure rate Ease of Operation +++ ++ Ease of Use Ease of Programming +++ + Ease of Use Real Time Overlays Yes No No No Improved reliability Integrate Chrom and UF/ DF Yes No No No Software 2 operations for the Sepragen Corporation price of one 43

Quanta. Sep Skids vs Competition Sepragen GE Pall Millipore Hardware Qsep 1000 Qsep 5000 Akta Process PK Series KPrime Series Benefits Product Range 6 3 3 3 Greater range Turndown Ratio of Pumps 100: 1 10: 1 30: 1 10: 1 Greater dynamic range Typical Flow Range 50 ml/min 5000 ml/min 6000 ml/min 100 ml/min 3000 ml/min 1500 ml/min Greater dynamic range Gradient (at 50% FS) 5 -95% 10 -90% 20 -80%% Better Chromatography Online Buffer Dilution Yes Yes No Save labor, space, time System Volume 750 ml ~2000 ml Better chromatography Footprint 22”x 27”x 46” 30”x 60”x 72” 30’x 60’x 72” 30”x 60”x 50” Increased Capacity Air Ejection Electronic Bubble trap Increased Reliability Leak Detection Yes No No No Increased Reliability Leak Sensor Yes No No No Increase reliability Easy to use software Yes No No No Easy training; usage and intervention Sepragen Corporation 44

Quanta. Sep vs Competition Sepragen Amersham Millipore Features Qsep 1000 Qsep 5000 3 -10 mm Bioprocess K Prime 40+ Benefits Inlet Valves 6+6, 4+4 4+4 Greater flexibility Columns 3, 1 1 1 Greater flexibility Fractions 10, 6 4 4 Greater flexibility Dual wavelength UV Yes Yes Greater flexibility Sepragen Corporation 45

Customer Quotes • Human Genome Sciences, Inc. "We have purchased several Quanta. Sep Systems and use them routinely in development and manufacturing. They have provided us with the capability to scale-up quickly and reliably. The robust design and compact size has enabled us to add productive capacity without waiting for additional construction. The Quanta. Sep software is easy to learn and use. Automated processing and report generation have enabled us to get more done with the same resources. Our quality and consistency have improved, as well. I would highly recommend the Quanta. Sep® family of products to anyone. " Mark Oelkuct, Purification Manager • Cohesion Technologies "At Cohesion we have used a Quanta. Sep 1000 as a workhorse in both manufacturing and process development. We are quite satisfied with its performance. In our hands the Quanta. Sep has been a reliable instrument that is also easy to use and maintain. The flow rate range of the 1000 matched out commercial requirement for one process and pilot scale for another. It fits well into smaller processing spaces (critical when working in Palo Alto). Again, we have been happy with our Quanta. Sep 1000 and would be willing to discuss our experience with others" Lou Sehl, Ph. D. , Senior Scientist and Team Leader Sepragen Corporation 46

Customer Quotes • Dade Behring "The efficiency of these products, in our hands, provides a significant advantage over other purification equipment we have evaluated for the following reasons: 1) The Quanta. Sep software is easy to use and train. 2) The Quanta. Sep allows us to reduce our standard direct hours, thus reduce our projected costs, by allowing a production chemist to perform multiple purification tasks simultaneously. This system eliminates the need to "baby-sit" all the individual purification steps. With reduced labor costs, the return on investment is easily less than two years for the Quanta. Seps. 3) This programmable system insures lot-to-lot consistency, resulting in consistent elution profiles vs. the built in variability with more manual operations. A critical advantage for a GMP production process. 4) And finally, one unique advantage of this system is its versatility. We can run both our small and large volume production (5 liter to 200 liter) runs with the change over feature. No other systems we have investigated have that volume flexibility. " Dennis Gould, Manager, Antibody Manufacturing Sepragen Corporation 47

Representative Quantasep Users for Quantaseps Domestic Customer Product International Country Company Product Siemen’s Diagnostic, Dyax Qsep 1000 LX Japan Asahi Kasei Qsep 1000 Neovatech Qsep 1000 s Germany Baxter Qsep 1000 Dow Pharma Qsep 1800/ 1000 Switzerland Lonza Qsep 1000 Lonza, Biogen- Idec Qsep 1000, 1800 Qsep 1000 India Cadilla Pharma Qsep 1000 Amgen, Roche Diagnostics Qsep 100 Belgium Henogen Qsep 1000 HGS Qsep 1000/ 1800 U. K. , Belgium GSK Qsep 1000, Qsep 40 Lpm Schering Plough Qsep 1000 s, 5000 China, Russia several Qsep 100/ Sepragen Corporation 1000 48

Summary § Reduce Project backlog § Improve product quality and reliability § Reduce Project backlog § Unattended clinical Production 24/7 NOW! § GMP documentation automatically § Easy scale-up 500 X § Improved process economic by 50% Sepragen Corporation 49

Quanta. Sep ® Additional Slides

Custom Explosion Proof Chromatography Skids • 10 -40 Liter per minute • Class 1, Div 1 or 2 • Stainless Steel construction • Fully drainable • Multi-wavelength detection • Diaphragm pumps Sepragen Corporation 51

Quantasep General Specifications Quantasep 100 Flowrate 1 – 100 m. L/min 10 – 1000 m. L/min Tubing ID (mm) 1. 59 (1/16”) 6. 35 (1/4”) Connections ¼. 28 thread ½” tri-clamp Hold-up volume 7 m. L 120 m. L Dimensions (Wx. Dx. H) 660 x 533 x 381(mm) (26"X 21" X 15") Un-crated weight (kg) 57 (125 lbs) 66 (145 lbs) Quantasep 1000 Sepragen Corporation 52

Sensor Specifications Sensor Range Accuracy Conductivity 0 -200 ms ± 2% p. H 1 -14 p. H ± 0. 3 UV (280 nm) 0 -2 Au ± 2% UV (260 nm) option Active Air Trap 0 -2 A-u ± 2% >5 mm (0. 20") 0. 6 x tubing ID Operating Pressure Leak 0 -7 bar (0 -100 psi) ± 0. 07 bar (± 1 psi) 5 m. L - Flow: Qsep 1000 Qsep 2000 1 -100 m. L/min 10 -1000 m. L/min 20 -2000 ml/min ± 3% Operating Temperature 4 - 30 o. C - Sepragen Corporation 53

Quantasep Flow cell Sepragen Corporation 54

Quantasep 100 - Inside Look Sepragen Corporation 55

Quantasep 1000 - Inside Look Sepragen Corporation 56

Flow cell and column & fraction manifolds for. Quantasep 1000 Sepragen Corporation 57

Multimode Detector Flow Cell p. H Ref p. H Probe UV Lamp Compact and Zero lag time detector helps minimize footprint Sepragen Corporation 58