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

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

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

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

Microcarriers: ATF2 System Setup 5 5

Microcarriers: ATF10 System Setup 6 6

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 40-50 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

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

Microcarriers: Seed Transfer Cell growth to maximum in batch or perfusion mode Media reduction at 1vv/hr for 30-40 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

Microcarriers: Media Exchange, Infection, Harvest Remove media at 1vv/hr for 40-50 mins Addition of new media Infect cells with virus Optional: Perfusion Harvest cells and debris at 1 vv/hr for 40-50 mins Diafiltrate for 10-20 minutes Optional: Clarification with ATF and 0.2u filter on holding tank ATF rate constant at 4-5x filtrate rate 10 10

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