1. © ChromaCon AG // www.chromacon.ch // Convert batch to MCSGP // ver. May 2013 1 Contichrom ® case study: How to convert a batch chromatography process into MCSGP

2. © ChromaCon AG // www.chromacon.ch // Convert batch to MCSGP // ver. May 2013 2 Questions How can I switch from a batch chromatography process to the MCSGP process? How long does it take to reach steady-state in the MCSGP process? What is the run time of an MCSGP process compared to a batch process Is the MCSGP process a continuous process?

3. © ChromaCon AG // www.chromacon.ch // Convert batch to MCSGP // ver. May 2013 3 Summary of the MCSGP process The process conversion of batch to MCSGP is easily done in three steps:

4. © ChromaCon AG // www.chromacon.ch // Convert batch to MCSGP // ver. May 2013 4 Starting with batch chromatography In a first step, a batch chromatography step needs to be developed. This includes: Stationary and mobile phase screening Loading and gradient optimization

5. © ChromaCon AG // www.chromacon.ch // Convert batch to MCSGP // ver. May 2013 5 Batch chromatogram features The batch process does not have to be optimized for yield and purity before conversion to MCSGP: 30%-70% yield at specified purity Duration of isocratic or gradient elution 10-40mins The product-containing regions are determined by offline HPLC analytics of collected fractions min 10-40mins UV-profile gradient Region of product in spec with 30%-70% yield Region with product containing impurities

6. © ChromaCon AG // www.chromacon.ch // Convert batch to MCSGP // ver. May 2013 6 Conversion into MCSGP The batch process is developed on the Contichrom ® equipment The corresponding batch chromatogram can then be uploaded using the wizard of the Contichrom ® operating software Using the wizard, the boundaries of the regions of pure product and product-containing impurities can be set by dragging bars to define these regions with the mouse as shown in the following animations

7. © ChromaCon AG // www.chromacon.ch // Convert batch to MCSGP // ver. May 2013 7 Step 1: retrieve chosen chromatogram of batch run from database Step 2: interactive definition of product range (red) and recycling fractions (blue): pull bars to define boundaries (dotted lines) Step 3: push button to convert batch to MCSGP process Automated conversion of batch to MCSGP method

8. © ChromaCon AG // www.chromacon.ch // Convert batch to MCSGP // ver. May 2013 8 Automated conversion of batch to MCSGP method 1. Load chromatogram of batch run Product peak

9. © ChromaCon AG // www.chromacon.ch // Convert batch to MCSGP // ver. May 2013 9 ANIMATION: Automated conversion of batch to MCSGP method 2. Interactive definition of product fraction

10. © ChromaCon AG // www.chromacon.ch // Convert batch to MCSGP // ver. May 2013 10 Easy conversion of batch to MCSGP chromatography side fractions stay in unit until pure waste pure product is withdrawn continuously

11. © ChromaCon AG // www.chromacon.ch // Convert batch to MCSGP // ver. May 2013 11 Flowsheet for the MCSGP process

12. © ChromaCon AG // www.chromacon.ch // Convert batch to MCSGP // ver. May 2013 12 How long does it take to reach steady-state in the MCSGP process? It takes 3-5 cycles to reach steady-state 1 st cycle UV detector signal of left column: 2 nd cycle 3 rd cycle 4 th cycle Steady-state reached UV detector signal of left column: 1 st cycle 2 nd cycle 3 rd /4 th cycle overlays of UV profiles of each cycle to accurately detect steady- state:

13. © ChromaCon AG // www.chromacon.ch // Convert batch to MCSGP // ver. May 2013 13 What is the run time of an MCSGP process compared to a batch process? The MCSGP process has typically the same transit time as a batch process Conventional scheme: Optimized scheme: Transit time:72 hrs 48 hrs 48 hrs (example)

14. © ChromaCon AG // www.chromacon.ch // Convert batch to MCSGP // ver. May 2013 14 Is the MCSGP process a continuous process? The MCSGP process is in most cases run only as a semi-continuous process, whereby the pure product is withdrawn at each cycle and the product-containing impurity fractions are recycled together with fresh feed. For a detailed representation, see the MCSGP process animation