1. An introduction to a novel filtration system
ATF-manufacturing Platform
Microcarrier Process Unit Operations
Vaccines
Gene Therapy
Stem Cells 1

2. Microcarrier Screen Filter Module
All USP Class VI materials (Polysulfone, Polyester, epoxy, 316L SS)
70um screen
Special flow path design to reduce fouling and attachment
Multiple low shear, rapid, separations, without settling:
Media exchanges, wash steps
Seed transfer
Harvest: cell debris & virus filtration from microcarriers 2

3. X X Maybe ? Microcarriers: Process Overview 3
Reactor seed train 5x volumes X X
Maybe ?
SIP: PBS & ucarriers
Wash ucarriers
Innoculate
Batch/perfusion growth
Trypsinization
Wash
Cell transfer
SIP: PBS & ucarriers
Wash ucarriers
Innoculate
Batch/perfusion growth
Trypsinization
Wash
Cell transfer
SIP: PBS & ucarriers
Wash ucarriers
Innoculate
Batch/perfusion growth
Media reduction
Infection
Media addition
Harvest
50+ flasks… 3 3

4. Microcarrier Processes: ATF System Advantages
Major advantage is liquid and microcarrier handling: ease and speed to remove or transfer media or cells
Rapid liquids removal without shear
Wash steps for carriers
Low shear cell/seed transfers
Rapid Harvest
Microcarriers do not need to settle
Cell ”health” improved, allowing higher productivity per cell
Media removal can be to a low volume
Simple to operate and scales to manufacturing
External system which allows easy exchange of filter if required 4 4

5. Microcarriers: ATF2 System Setup5 5

6. Microcarriers: ATF10 System Setup6 6

7. Microcarriers: Preparation & Washing
Reactor filled with carriers and PBS
Reactor is SIP’d
ATF is autoclaved
Steamed connection made
PBS removed at 1vv/hr for mins
Addition of media
Diafiltrate for 5 minutes
Close harvest, media added to working volume
Innoculate with cells
ATF rate constant
at 4-5x filtrate rate 7 7

8. Microcarriers: Perfusion
Perfusion carried out at small or large scale
Rates at 0.5vv/day to 4vv/day
ATF rate at 30+ higher than filtrate rate 8 8

9. Microcarriers: Seed Transfer
Cell growth to maximum in batch or perfusion mode
Media reduction at 1vv/hr for mins
Trypsinise cells
Harvest cells through ATF leaving behind carriers
Diafiltrate with new media, to capture maximum number of cells
Cells transferred directly to next reactor
ATF rate at 3-5x filtrate rate 9 9

10. Microcarriers: Media Exchange, Infection, Harvest
Remove media at 1vv/hr for mins
Addition of new media
Infect cells with virus
Optional: Perfusion
Harvest cells and debris at 1 vv/hr for mins
Diafiltrate for minutes
Optional: Clarification with ATF and 0.2u filter on holding tank
ATF rate constant
at 4-5x filtrate rate 10 10

11. Microcarrier-free Harvest
Adherent Cells: Combined DSP
Buffer, Wash
Vaccine Process – Adherent Cells
Ionic Solution
(hc-dna precipitation)
70u filtered stream
0.2u or 0.5u filtered stream
Clarified Harvest
Microcarrier-free Harvest
Production reactor