Automated manufacture of T-cell immunotherapies using gamma retroviral transduction. Lee Markwick, PhD. AMC, Manchester March 2018

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1 Automated manufacture of T-cell immunotherapies using gamma retroviral transduction Lee Markwick, PhD AMC, Manchester March 2018

2 Automated generation of CAR T cells Starting material d0 T cell enrichment and activation d1 Transduction d2-10 Expansion d10-14 Final product (max of 14 days) 8th March 2018 Automated manufacture of T-cell immunotherapies - Lee Markwick Ph. D. Page 2

3 CliniMACS Prodigy TCT Process Cell selection Cell activation Cell transduction Cell expansion Final formulation IPC/QC CliniMACS Buffer MACS GMP TransAct GMP Lentiviral Vector MACS GMP TexMACS CliniMACS Reagents: CD4 / CD8, CD62L MACS GMP TexMACS CliniMACS Prodigy TS 520 CliniMACS Prodigy TCT Process for RV and LV MACS GMP IL-2, IL-7, IL-15, IL-21 flow reagents and REA antibodies 7 color immunophenotyping kit MACSPlex Cytokine Kit Rapid Cytokine Inspector Kit 8th March 2018 Automated manufacture of T-cell immunotherapies - Lee Markwick Ph. D. Page 3

4 TCT process - robust and reproducible with pts material Cell selection Cell activation Cell transduction Cell expansion Final formulation Robust expansion of healthy material (n=12) and pts material (n=6) Good viability of healthy material (n=12) and pts material (n=6) Patient material T cells CAR + T cells Melanoma 2.35E9 1.38E9 Lymphoma 3.68E9 1.54E9 Lymphoma 3.31E9 - Lymphoma frozen 4.90E9 1.09E9 Lymphoma frozen 3.20E9 0.72E9 E.g. dose: 10 7 CAR + T cells / kg (130kg patient) 8th March 2018 Automated manufacture of T-cell immunotherapies - Lee Markwick Ph. D. Page 4

5 TCT process - robust and reproducible in serum free conditions Cell selection Cell activation Cell transduction Cell expansion Final formulation 8th March 2018 Automated manufacture of T-cell immunotherapies - Lee Markwick Ph. D. Page 5

6 Volume reduction Addition RV + enhancer Spin 400g, 32 C Culture wash Start shaking New Activities to enable TCT for retroviral vector TCT Retro Enrichment Volume chamber Activation in 70 ml day h incubation time 8th March 2018 Automated manufacture of T-cell immunotherapies - Lee Markwick Ph. D. Page 6

7 Retroviral transduction Cell selection Cell activation Cell transduction Cell expansion Final formulation IPC/QC Vectofusin-1 Transduction enhancer Fully synthetic, non toxic, soluble peptide No pre-treatment/coating needed - added directly to viral supernatant! Straight forward and simple protocol Available in research and GMP grade 8th March 2018 Automated manufacture of T-cell immunotherapies - Lee Markwick Ph. D. Page 7

8 Retroviral Transduction Cell selection Cell activation Cell transduction Cell expansion Final formulation 8th March 2018 Automated manufacture of T-cell immunotherapies - Lee Markwick Ph. D. Page 8

9 Cellular composition of final product Cell selection Cell activation Cell transduction Cell expansion Final formulation 8th March 2018 Automated manufacture of T-cell immunotherapies - Lee Markwick Ph. D. Page 9

10 Functionality: Killing of CD20+ JeKo-1 cells Cytokine secretion profiles Healthy donor Cytotoxicity Healthy donor Patient material Patient material Automated manufacture of T-cell immunotherapies - Lee Markwick Ph. D. 8th March 2018 Page 10

11 Robust manufacturing of polyfunctional gene-engineered T cells: In vivo mouse model 8th March 2018 Automated manufacture of T-cell immunotherapies - Lee Markwick Ph. D. Page 11

12 Cell Manufacturing Centres 8th March 2018 Automated manufacture of T-cell immunotherapies - Lee Markwick Ph. D. Page 12

13 Integral Single Unit Manufacturing Parallel Manufacturing. Instantly scalable More devices, more products Simplicity at Scale. One process, one device Complexity Minimisation: Process steps Connections Device upkeep & maintenance 8th March 2018 Automated manufacture of T-cell immunotherapies - Lee Markwick Ph. D. Page 13

14 Facility Considerations (Layouts for 2000 patients p.a.) David James Cell Gene Therapy Insights 2017;3(4). 8th March 2018 Automated manufacture of T-cell immunotherapies - Lee Markwick Ph. D. Page 14

15 CliniMACS Prodigy in clinical trial and licensed commercial product 8th March 2018 Automated manufacture of T-cell immunotherapies - Lee Markwick Ph. D. Page 15

16 Miltenyi Biotec manufacturing suite 8th March 2018 Automated manufacture of T-cell immunotherapies - Lee Markwick Ph. D. Page 16

17 Automation & Operation March 13, 2018 Jason Jones, Miltenyi Biotec Ltd Page 17

18 Parallel manufacturing of cell and gene therapy products March 13, 2018 Page 18

19 Not just CAR T-cells Blood Leukapheresis Cell selection Cell activation Cell transduction Cell expansion Cell formulation Patient Donor / patient MoDC CAR T cells Treg cells Antigen-specific T cells Gene-modified stem cells TCR T cells NK cells MSCs 8th March 2018 Automated manufacture of T-cell immunotherapies - Lee Markwick Ph. D. Page 19

20 Your trusted partner for cell and gene therapy 8th March 2018 Automated manufacture of T-cell immunotherapies - Lee Markwick Ph. D. Page 20

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