Cell Characterization: Developing Assays to Address the Biological Parameters of Identity and Potency

Transcription

1 Cell Characterization: Developing Assays to Address the Biological Parameters of Identity and Potency Jessica Carmen Ph.D and Jon Rowley Ph.D., LBS-TCS, May 2011

2 Disclaimer Certain matters discussed in this presentation may constitute forward-looking statements. These statements are based on current expectations and estimates of Lonza Group Ltd, although Lonza Group Ltd can give no assurance that these expectations and estimates will be achieved. The actual results may differ materially in the future from the forward-looking statements included in this presentation due to various factors. Furthermore, Lonza Group Ltd has no obligation to update the statements contained in this presentation. slide 2

3 Lonza Cell Therapy Services Tissue acquisition Media formulation Process development and optimization Veterinary services Assay development and validation Cell banking cgmp manufacturing of both autologous and allogeneic therapies Product testing and release Regulatory filing support Packaging Distribution slide 3

4 Challenges for Scale Up Master & Working Cell Banks Comparability Risk Tissue Acquisition Primary Cell Isolation Cell Culture Harvest Volume Reduction Storage & Inventory Formulate & Process Washing Fill Additional population doublings boost lot size Culture technologies Vessels Growth surfaces Media Testing Serum & reduction / elimination Shipping Release Logistics efficiencies via feed strategies Cryopreservation End User Handling & Delivery Lonza Strength: Innovation in Large-Scale Process Development slide 4

5 Cell Therapy Critical Quality Attributes Identity to confirm that the type of cells in the final product container are the intended ones Potency to confirm that the cell product possesses the appropriate set of biological functions that are relevant to treating the intended clinical indication Purity to confirm that the product is free from harmful contaminants (unwanted cell types) and process residuals (animal serum and trypsin) Safety to confirm that the product poses no risk to a patient due to contamination with microbes, endotoxin, or adventitious agents and, if appropriate, the tumorigenicity profile of the final cell population slide 5

6 Cell Therapy Critical Quality Attributes Identity to confirm that the type of cells in the final product container are the intended ones Potency to confirm that the cell product possesses the appropriate set of biological functions that are relevant to treating the intended clinical indication Purity to confirm that the product is free from harmful contaminants (unwanted cell types) and process residuals (animal serum and trypsin) Safety to confirm that the product poses no risk to a patient due to contamination with microbes, endotoxin, or adventitious agents and, if appropriate, the tumorigenicity profile of the final cell population slide 6

7 The Identity Crisis Manufacturing of cell therapy products for early-stage clinical trials often rely on identity (FACS) for comparability Identity alone is not sufficient to ensure biological functionality through process development We will show examples of process development activities in which identity (flow cytometry data) is comparable while potency measures are changing Extended culture Serum elimination slide 7

8 Extended Culture

9 MSCs Grown to 10 Passages in 10-layer Cell Factories Human MSCs from 1 donor were grown to 80% confluence in 3 10-layer cell factories At passage 1, 3, 6, and 10 cells were sampled for flow cytometry, cytokine secretion, and IDO activity Passage numbers were converted to population doublings Passage PDL slide 9

10 Consistent Surface Marker Expression slide 10

11 Significant Changes in Cytokine Secretion Through Time slide 11

12 IDO Activity Declines with Increasing Population Doublings slide 12

13 Extended Culture It is important to test the limits of cell expansion in order to maximize product lot size without compromising biological function slide 13

14 Serum Elimination

15 MSCs Grown in T-flasks Using Serum- Containing Versus Chemically Defined Media MSCs from 1 donor were grown in the following medias for three passages: Lonza MSCGM (10% serum) Lonza MSCGM-CD (chemically defined) Competitor s serum-free media with proprietary attachment substrate At final harvest (cumulative P6) cells were sampled for flow cytometery, cytokine secretion, and IDO activity slide 15

16 Consistent Surface Marker Expression slide 16

17 Significant Differences in Cytokine Secretion slide 17

18 IDO Activity Differs with Different Chemically Defined Medias slide 18

19 Chemically Defined Media Screen multiple media when removing animal products When efforts are made to reduce/eliminate animal origin products in cell therapy manufacturing, only looking at identity is not sufficient to show functional changes in your cells and you could see clinical efficacy changes slide 19

20 Cell Characterization and Process Development Altered biological function is acceptable as long as the new activity levels are taken into account prior to release specifications being set Release specifications are set through iterative testing during the manufacture of clinical trial materials Cell characterization and process development activities occurring early in the manufacturing process will confirm that the potency assay specifications match the product produced with the improved process slide 20

21 Acknowledgements Lonza Therapeutic Cell Solutions R&D Elisabeth Misleh Lonza Therapeutic Cell Solutions PD Wayne Hildebrand Heather Dernoeden April Birch Lonza Bioservices Ashley Rimshaw Joseph Finny Ph.D. slide 21

22 Thank you for your attention