an introduction to a novel filtration system atf-manufacturing platform microcarrier process unit...
TRANSCRIPT
![Page 1: An introduction to a novel filtration system ATF-manufacturing Platform Microcarrier Process Unit Operations Vaccines Gene Therapy Stem Cells](https://reader035.vdocuments.net/reader035/viewer/2022081603/56649c7c5503460f94930f02/html5/thumbnails/1.jpg)
An introduction to a novel filtration system
ATF-manufacturing PlatformMicrocarrier Process Unit Operations
VaccinesGene TherapyStem Cells
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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
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Microcarriers: Process Overview
SIP: PBS & ucarriers
Wash ucarriers
Innoculate
Batch/perfusion growth
Trypsinization
Wash
Cell transfer
Reactor seed train 5x volumes
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…
X XMaybe ?
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• 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
Microcarrier Processes: ATF System Advantages
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Microcarriers: ATF2 System Setup
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Microcarriers: ATF10 System Setup
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1. Reactor filled with carriers and PBS
2. Reactor is SIP’d
3. ATF is autoclaved
4. Steamed connection made
1. PBS removed at 1vv/hr for 40-50 mins
2. Addition of media
3. Diafiltrate for 5 minutes
4. Close harvest, media added to working volume
5. Innoculate with cells
ATF rate constant
at 4-5x filtrate rate
Microcarriers: Preparation & Washing
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1. Perfusion carried out at small or large scale
2. Rates at 0.5vv/day to 4vv/day
3. ATF rate at 30+ higher than filtrate rate
Microcarriers: Perfusion
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1. Cell growth to maximum in batch or perfusion mode
2. Media reduction at 1vv/hr for 30-40 mins
3. Trypsinise cells
4. Harvest cells through ATF leaving behind carriers
5. Diafiltrate with new media, to capture maximum number of cells
6. Cells transferred directly to next reactor
7. ATF rate at 3-5x filtrate rate
Microcarriers: Seed Transfer
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1. Remove media at 1vv/hr for 40-50 mins
2. Addition of new media
3. Infect cells with virus
4. Optional: Perfusion
1. Harvest cells and debris at 1 vv/hr for 40-50 mins
2. Diafiltrate for 10-20 minutes
3. Optional: Clarification with ATF and 0.2u filter on holding tank
ATF rate constant
at 4-5x filtrate rate
Microcarriers: Media Exchange, Infection, Harvest
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Production reactor
0.2u or 0.5u filtered stream
Clarified Harvest
Adherent Cells: Combined DSP
Vaccine Process – Adherent CellsBuffer, Wash
70u filtered stream
Microcarrier-free Harvest
Ionic Solution(hc-dna precipitation)