Ancillary Materials in Cell Therapy Product Development
Transcript of Ancillary Materials in Cell Therapy Product Development
Ancillary Materials in
Cell Therapy
Product Development
Nicole M. Provost, PhD Independent Consultant
Member of USP BB2 Expert Committee
Ancillary Material
Topics to be Covered
Common Sense Approaches
Definitions and Abbreviations
CMO Advantages and Disadvantages
Risk-based Approaches
“Big 3” Questions to Address
Relevant Examples
Take-home Messages
(Intuitively Obvious )
Common Sense Approaches
Figure out what you know, don’t know
Don’t ignore problems
Make plans
Be ready to change your plans
Definitions and Abbreviations
Ancillary Material (“AM”)
◦ Material used and removed during manufacture of a cell or tissue product
Supplier
◦ “Off-the-shelf” manufacturer of AM
Contract Manufacturing Organization (“CMO”)
◦ Custom manufacturer of AM
Product Development (“PD”)
Research and Development (“R&D”)
CMO Advantages
AM Manufactured by CMO
◦ No Manufacturing and Quality personnel to
hire and oversee
◦ Reduced need for additional facilities
◦ Reduced need for additional headcount
Supply Chain Managed by CMO
◦ Fewer suppliers to oversee
◦ Shipping and storage managed by CMO
◦ Multiple suppliers = more flexibility
CMO Disadvantages
AM Manufactured by CMO
◦ Ultimately, CMO issues become yours
◦ Be prepared to troubleshoot
◦ Designate a contact person on your side
Supply Chain Managed by CMO
◦ Sourcing is out of your control
◦ Shipping and storage issues can disrupt your
own manufacturing and R&D
◦ Multiple suppliers may = more variability
Risk-based Approaches to AMs
in a Nutshell
Perform Criticality Assessments
Identify the major risks to cell product: purity, consistency, supply, stability, etc.
Determine AM & product failure mode(s)
Define product/patient impacts
Mitigate or eliminate the major risks
Decide which headaches you own, and which you share with the CMO
Build trust with your CMO
The “Big 3” Questions for PD
What’s in the Ancillary Material?
How does the Ancillary Material affect
the cell product?
• Can the CMO make enough Ancillary
Material?
What’s in the Ancillary Material?
Is it a complex mixture?
◦ Classic examples: culture media and serum
Identify & characterize active component(s)
◦ Sometimes the impurities are active!
◦ Conduct fractionation and add-back experiments
◦ Relative component ratios may be important
Identify major impurities
◦ Common examples: fragments, endotoxin, DNA
How variable are the batches?
◦ Quantitation and characterization assays critical
How Does the AM Affect
the Cell Product?
Is the AM essential to cell product quality?
◦ Damage or omit the AM to assess its criticality
◦ Determine concentration effects, stability in use
◦ Assess many batches or sources
Do impurities affect cell product quality?
◦ Add-back experiments with known impurities
◦ Determine safety limits
Are both AM and impurities removed during cell product manufacturing?
◦ Both can be washed out, taken up by cells, or adsorbed onto production equipment
Can the CMO Make Enough AM?
How much AM do you really need?
◦ Perform titration studies early on
◦ Aim for a broad concentration “sweet spot”
How stable is the AM?
◦ Determine stability under storage, stress, and manufacturing conditions
Can the CMO scale up production?
◦ Look for a back-up supplier
Are you a major customer?
◦ This can be a mixed blessing for everyone
Relevant Examples
DMSO cryoprotectant, and plasticizers
Culture media formulations
Endotoxin impurities
Ancillary Material breakdown byproducts
Scale-up headaches
Plasticizers Leach
from Plastic into Cells
Inoue et al. (2005) Clinica Chimica Acta 358, pp. 159-166
DMSO Cryoprotectant,
and Plasticizers
DMSO increases plasticizer leaching
Check your DMSO source
DMSO can affect the integrity of
equipment, seals and filters
Plasticizers can affect cells, even if DMSO
is washed out
Conduct plasticizer add-back experiments
to estimate risks
Culture Media: Points to Consider
Natural, synthetic, recombinant
◦ Albumin, ferritin, lipids, “secret sauce”
◦ Scale-up, cost, GMP availability issues
Powder vs. liquid formulations
◦ Impacts on stability, cost, logistics, and risk
◦ Switching between liquid & powder is difficult
Lipids can be critical
◦ Solubility issues (especially in powders)
◦ Multiple lipid species, variable amounts
pH indicator – do you really need it?
◦ May be taken up by cell membranes
◦ Useful marker for tracking wash efficiency
Endotoxins in Ancillary Materials
Present in multiple components
◦ Additive effects during manufacturing
◦ Cleanliness indicator for AM manufacturing
“Sticky”, not easily washed away
◦ Added risk for multi-use equipment
Pleiotropic effects on cell products
◦ Inflammatory and cytotoxic effects
◦ Can initiate cytokine cascades
Establish limits for patient safety
◦ Regulatory and safety requirements
◦ Conduct spiking experiments
AM Breakdown Byproducts
Cell product manufacturing conditions can
degrade AMs
◦ Temperature, agitation, pH, oxidation
◦ Proteolytic enzymes from damaged cells
◦ Incubate with conditioned medium
AM Fragments may be more active
◦ Chemical adducts, fusion proteins, etc.
De-glycosylation may affect bioactivity
Characterization assays are critical
AM Scale-up Headaches
Check with your AM supplier
◦ Batch sizes, manufacturing frequency, shelf life, storage conditions, constraints
◦ Impurity profiles can change with scale
Scale-up potential may be limited
◦ Expensive or unstable ingredients
◦ Equipment and facility constraints
◦ Not enough demand
◦ Environmental restrictions
Scale up early in cell product life cycle
◦ Phase 3 surprises are always unwelcome
Take-home Messages
Characterize the AM and cell product early and often in development
◦ Invest in characterization, quantitation, and activity assays early on
◦ Analyze failure modes and stability profiles
◦ Assess and mitigate risks
◦ Measure residuals in final cell product
Work with the AM supplier
◦ Minimize risks
◦ Maximize yields