1. An introduction to a novel filtration system ATF-manufacturing Platform Upstream Unit Operations ATF-cellcultivation Concentrated Perfusion Concentrated Fed-Batch Hybrid Processes

2. 2 Capacity cannot be calculated solely by bioreactor size 10kg500g1000L 3kg150g300L 1kg50g100L 20 Runs/YrPer Run *Fed-Batch 150kg15kg500g1000L 45kg4.5kg150g300L 15kg1.5kg50g100L 10 Runs/Yr30 Day RunPer Day *Perfusion * 500 mg/L post purification yield 15x more product in the same time! Avid Bioservices Facilities

3. Why use Perfusion in Research, even if not later... Perfusion produces more material in a shorter time period –Do you have time to make a new cell line? –Do you have time to develop a special media? Pre-clinical material can be produced faster with less development effort A low producing cell line can be used to produce enough material in a shorter time for a phase 1 trial Smaller bioreactors are required at all stages of PD and Manufacturing - saving significant capital cost One continuous culture experiment can perform multiple metabolic studies to understand clone behavior in different design spaces, significantly faster than in batch or fed-batch Upstream development times may be reduced due to simpler feeding

4. Perfusion and Concentrated Perfusion Perfusion is dead - Long Live Concentrated Perfusion !! What is Concentrated Perfusion? –Only possible with the ATF –Ultra-high viable cell densities, in the region of 60-150m / ml –Requires 1-3 vv / day media (not more) –Same simple process control as perfusion & CFB – but higher productivity When should perfusion be used and not Concentrated Perfusion? –When the product quality is affected by cell concentration, or if protein expression depends on a dilute toxic species

5. Concentrated Perfusion “Standard” perfusion application but with the ATF System –high cell concentration (60-100m/ml), 30-60 days Straight-forward feeding control, removal of metabolites & proteins Continuous filtered harvests, often pooled (every 3-6 days) for easier DSP –~0.1g/L/day per 10m cells /ml for an average cell line (10pg/cell/day at 2vvd) MF filter retains cells inside reactor while molecules filter through Filtrate pump runs at 0.5-3 vv / day or as required by cell growth ATF rate constant at minimum ~0.05-0.1 vv / min Feeding controlled by level probe

6. Graph taken from Genetic Engineering News October 2007 CHO Perfusion Results