Transcript Présentation PowerPoint - ISPE-CaSA

Controls and
containment in
plant-based
vaccine
manufacturing
Todd Talarico
Vice President of Process
Development and Clinical
Manufacturing
Objective
 Introduce Plant Based Biologics
 Present Controls used for Manufacturing based upon:
– Health Canada : Guidance Document – Submission Information for Human-Use
Biological Drugs Derived from Plant Molecular Farming (PMF)
– FDA: Draft Guidance for Industry: Drugs, Biologics and Medical Devices Derived
from Bioengineered Plants for Use in Humans and Animals
– ICH Q7 - Good Manufacturing Practice Guide for Active Pharmaceutical
Ingredients
– ICH Q5 series – Quality of Biotechnological Products
– EMEA – Guideline on the Quality of Biological Active Substances Produced by
Stable Transgene Expression in Higher Plants
• Guideline on Good Agricultural and Collection Practice (GACP)
 Focus on the process first and then containment
– Familiar Principles
– Principles that may be more unique to plant derived biotherapeutics and
vaccines
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Medicago Overview
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Focus
Vaccines & Therapeutic Proteins
Manufacturing technology
Transient expression in plants
Vaccine technology
Virus-like particles
Employees
~300 Expected to grow to 500 by 2019 (commercial)
Success story
Innovative Vaccine Technology
Potential advantages over current vaccine
technologies (efficacy, cross protection)
Pandemic Supply advantages
Headquarters, laboratories
& cGMP facilities
Quebec City, CANADA
Research Triangle Park, NC, USA
Corporate structure
Private company
Mitsubishi Tanabe Pharma Corporation (majority
shareholder), PMI
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Plant Platforms for the Production of Biologics
• Plant cells (bioreactor)
• Whole plants
• Expression systems
• Transgenic plants with nuclear transformation
• Transient expression
• Induced by plant specific bacterium or virus
polynucleotide transfer
• Advantages of transient expression system
» No propagation through seed stock
» Genetic stability of expression
• Growth systems
• Indoors (Greenhouses, Growth Chambers)
• Outdoors with restrictions
• Expression
• Indoors (Greenhouses, Chambers) or Outdoors
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Medicago’s Technology
• Plants used for production are not genetically
modified and grown in a greenhouse
• Use a genetically engineered Agrobacterium to
drive expression of a protein in the leaves of
“tobacco” plants
• In the case of the Influenza vaccine, the plants
produce virus like particles (VLP) containing
the HA protein imbedded in a lipid bilayer when
incubated under controlled conditions
• The VLP are extracted from the plant tissue
• The VLP are purified in a “traditional”
bioprocessing scheme to produce a sterile
product, suitable for injection
Medicago’s Bioreactor
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Key Aspects of Medicago’s Process Control
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• Stock maintenance
– Plant seed stock maintenance system
– Agrobacterium MCB/WCB maintenance
• Upstream
• Greenhouse
• Stock to harvest
• Before infiltration
– Defined biological starting material
• After infiltration (protein expression)
• Downstream
– After extraction
– Purification, Formulation and Filling
 Facility Control
 Equipment Control
 Personnel Control
 Raw Material Control
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MEDICAGO QUADRIVALENT VLP INFLUENZA VACCINE CONTROL STRATEGY
Control of Materials
Manufacturing Process
Control
Characterization and specification
of Biological Starting Materials
- Plants
- Agrobacterium
- Enzymes
Procedural Control
- CP-SOP & Site SOP
- Batch Records
- Risk Assessment and Mitigation
Drug Substance Control
- SOPs
- Batch Records
- Acceptable ranges for standard
operations via Process validation
- Specifications
Process Characterization
- Data Mining
- clinical product history campaign
summary reports
Drug Product Control
- SOPs
- Batch Records
- Acceptable ranges for standard
operations via Process validation
- Specifications
Process Parameters Controls
- Identification of Critical process
parameters (CPP)
- Rationale for CPP
- CPP limits via process validation
Process Monitoring
- Evaluation of selected attributes
and/or parameters
- Trend process performance in
design space
- Include limits to evaluate data
trends
- Adjust frequency of periodic
monitoring based on trends
The Monovalent VLP DS Process
Control Strategy was developed to
minimize the variability caused by
input parameters in accordance with
the principles found in the CMCVaccines Working Group document,
A-VAX: Applying Quality by Design to
Vaccines, issued in May 2012.
Analytical/Testing
Control
In-Process Testing
Ensure selected manufacturing
operations are performing
satisfactorily to achieve the
intended product Quality
Release Testing
Final Tests associated with
acceptance criteria performed on
a set of quality attributes to
confirm the quality of drug
substance for forward processing
and drug product for distribution
Characterization
Testing of certain attributes
outside of lot release testing for
the purposes of demonstration of
comparability
Monitoring
-Testing of selected attributes To
trend product quality
- For enhanced confidence in the
normal distribution of an attribute
Matching Process and Analytical controls
FOR
Identification of Critical Quality Attributes and Critical Process
parameters
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Raw Material
Utilities
Personnel
Production areas
etc
Guidance Documents that support the plant platform
Health Canada : Guidance Document – Plant Molecular
Farming (PMF) Applications: Plant-Derived Biological
Drugs for Human Use (2014)
FDA: Draft Guidance for Industry: Drugs, Biologics and
Medical Devices Derived from Bioengineered Plants for
Use in Humans and Animals (2002)
ICH Q7 - Good Manufacturing Practice Guide for Active
Pharmaceutical Ingredients
ICH Q5 series – Quality of Biotechnological Products
EMEA – Guideline on the Quality of Biological Active
Substances Produced by Stable Transgene Expression in
Higher Plants (2008)
– Guideline on Good Agricultural and Collection Practice
(GACP)
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Quality Requirements for Plant Derived Biologics
Meet ‘the same quality standards as drugs produced by
conventional platforms’
‘PMF-derived drugs are subject to the same general
submission requirements…as other biological drugs’
Address ‘risks unique to PMF production platforms,
particularly in the pre-harvest and harvest stages …’
– Concern about control of plants in a field
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Stock Maintenance
 All starting material must be defined
– Origin and maintenance
 Agrobacterium stock
– Managed through the use of Master and Working Cell Banks
– Based on principles defined in ICHQ5
 Plant seed stock maintenance
– Nicotiana benthamiana
• Self germinating - homogeneity
• Propogated during winter in Quebec (to prevent foreign pollen introduction)
– Parenteral Seed Stock
• Regular (5 year) regeneration with cumulative stability being monitored
– Annual Seed Stock
• Produced annually from parental stock
• Aliquoted and stored in independent locations
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Quality System (‘GMP-like’ controls) – to produce a welldefined biological starting material
–
–
–
–
–
–
Staff qualification and training
Written procedures
Incoming material specification
Batch records
Biological material specifications for plants and harvested material
Trending, including monitoring of production consistency, environment
• Characterization such as fiber content, Mycotoxins, Mold + yeast, plant virus,
bacterial flora, substrate pH, substrate conductivity
– Identifying key control parameters
• Product safety first
• Parameters that have an impact on product attributes and yield
– Other traditional oversight - Risk Assessment, Audits, Change Control
– Equipment controls
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Greenhouse (Upstream)
Controlled process that results in a defined biological
starting material
The greenhouse environment is designated as a
“controlled, not classified” environment
Quality system with QA and QC components
– ‘GMP-like’ principles
– Not all pharmaceutical GMP regulations can apply
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Production of Plants
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Medicago’s Controls on Plants
 All operations covered by production batch records for generation of
the critical raw material
 No herbicides, pesticides or fungicides
 Specifications for input raw materials
– Peat, fertilizer, water
 Control of entry
– Personnel, filtered air (not HEPA), insect monitoring, rodent exclusion
and monitoring
 Criteria for acceptance as plants proceed from seeding to infiltration
 Routine checks for plant viruses and organisms that could produce
mycotoxins
 No documented cases of human viruses infecting and replicating in
plants or plant viruses replicating in humans
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Plant Control
 Controls are in place to assure plants provide proper protein expression
 The “working operating space” for many variables has been demonstrated to
be large which makes for a robust production system
– 20 to 50 gm plants have been used to produce the influenza vaccine with
no negative impact to yield or CQA
 Other variables have more impact on protein expression
– Flower stage is important to control
 The plant has been mapped to understand which tissues produce the most
product
 Kinetics of expression are determined for each strain
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Plant Certificate of Analysis
Test
Document et version /
Critère d’acceptation
Document number and version
Acceptance criteria
W (Largeur de la plus grande
feuille / Width of the largest
Résultat / Result
≥7
leaf)
S (Nombre de tiges
secondaires / Number of
secondary stems)
F (Stade de dévelopement
de fleurs et graines / Flower
≥2
≤6
and seed stage)
Uniformité du lot global /
≥ 80%
Overall batch uniformity
TEST
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DOCUMENT ET VERSION /
DOCUMENT
NUMBER AND VERSION
RÉSULTAT / RESULT
Les plantes utilisées ne présentent pas de
symptômes de maladie / Plants used are free
Oui / Yes
from disease symptoms
Non / No
Les plantes utilisées ne montrent pas une
présence élevée d’insectes ravageurs / Plants
Oui / Yes
used are free from an elevated pest pressure
Non / No
Le lot de plante est conforme /
Oui / Yes
The plant lot is compliant
Non / No
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Downstream
 After extraction – purification is similar to other biological products
 Plant specific concerns – real or perceived
– Impurities from plant manufacturing process
• Fibres, lipids, host cell proteins
• Plant secondary metabolites
– Risk assessment
• Consider as well the potential allergenic potential
• Define their nature
• Reduce to safe levels
– Plant glycans (part of the glycoproteins)
• Potential diffierences in glycan structure
• Analysis and risk assessment
– No evidence they are immunogenic
– Could be beneficial to process compared with mammalian cell glycans
• Product and use specific (vaccine vs long term therapeutic)
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Process Validation (PPQ)
 Master validation plan outlines methods
 Data mining performed on 3 production process generations
 Data from > 20 than runs used to set CPP ranges based on CQA of
vaccine
 Secondary dataset for current process compared back to original
data mine
 These data will be used to set the acceptable ranges for the
consistency lots used for Phase 3 manufacture
 Agreement on proposal to the Agency for consistency lots
 Obtained buy in for number of lots per strain and number of runs to
generate drug substance
 Three fills planned with CMO to validate filling process
 Matrixed based on number of units filled
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CPPs for the Production Process
 5 CPP around filtration operations
 3 CPP around chromatography
 1 CPP for virus inactivation
 Quality mandated CPP for filtration (2) and virus inactivation (1)
– for example – Sterility
 Moving forward some CPP may be eliminated by incorporating PAT
or reducing risk
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Containment – Material and Process
 Clarification and downstream operations are similar to traditional
bioprocessing
– Use of disposables
– Where stainless steel is used, cleaning validation is traditional
• Look for plant based compounds (nicotine, chlorophyll, phenolics)
– Closed systems as we approach drug substance
• Single use bag technology
– VLPs pass through a 0.2 µ filter
– Facility design utilizes typical classifications
 Greenhouse and VLP (or protein) extraction are the areas that are
dissimilar to traditional bioreactor based biopharmaceutical
operations
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Process Challenges from a “Contaminant” Perspective
 A greenhouse is not an aseptic environment
 Efficient operations require recycling of materials
– Water, fertilizer
 Biological raw materials
– No animal derived components
– Many not be “low bioburden or sterile”
 Agrobacterium is Gram negative
– Endotoxin (but of low pyrogenicity)
 Human interaction in an open system with critical raw materials
– Plants
 A VLP is much like a virus from a surface biochemical point of view
– Hard to get a process to have high levels of viral removal/inactivation
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Bioburden Clearance in the Process
 Centrifugation and microfiltration early in the process reduce
bioburden
 Sterilizing grade filter removes Agrobacterium early in the process
and reduces all bioburden prior to process intermediate entrance into
downstream operations
 Additional microfiltration protects chromatography columns and
reduces any bioburden load
 Sterile filtration prior to fill of bulk drug substance and again prior to
fill to syringes or vials
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Viral Risk
 Plants are considered as a low risk for introduction of adventitious
agents
– Human viruses have not been shown to replicate in plants
– Plant viruses are not known to infect human cells
– Plants act as a barrier to virus introduction into the process
 No animal derived components used in the process
 Largest risk is introduction of an adventitious agent into the process
by human, rodent or insect.
 Regulators worry about new threats emerging based upon past
experience in biological processes
 Plant based production system presents a low risk of AA introduction
when the proper controls are in place
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Viral Clearance
 Many negotiations with the Agency to move from safety testing to
inactivation
 Inclusion of a UVC treatment unit operation in the process
 Development work performed to identify the optimal dose to
inactivate a variety of viruses with minimal impact to the product
 Added bonus of spiroplasma and mycoplasma inactivation
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UVC Development
 5 Viruses used to assess system effectiveness
 Wide range of doses tested
 Looked at inactivation efficiency and impact to product
 Data mining from process to assess control limits and risk
Virus
200
>7.64 ± 0.06
>5.93 ± 0.05
>5.44 ± 0.04
3.17 ± 0.04
MVM
BVDV
PRV
17.0
20
15.5
H3
15
10
5
0
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8.4
9.3
350
>7.70 ± 0.06
>7.27 ± 0.05
>5.55 ± 0.09
3.78 ± 0.05
UVC Flow Rates L/hr
UVC Lamp Intensity mA
20
Dose J/m2
270
>7.61 ± 0.06
>7.26 ± 0.05
>5.44 ± 0.04
3.45 ± 0.04
B Bris
15
10
B Mass
H1
5
0
A254 Values
19.0
17.7
17.5 18.4
5
H3
4
B Bris
3
B Mass
2
H1
1
3.4
2.9
H3
1.3
1.0
B Bris
B Mass
H1
0
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UVC Viral Inactivation
 Process validation completed
–
–
–
–
Scaled down model equivalent to manufacturing process
Rationale documented to support chosen studies (Risk based)
Used the development data to set boundries for validation study
Duplicate runs with three viruses using two HA types at a single UVC
dose
– Robustness studies to assess processing and equipment extremes at
chosen UVC dose
– Re-use of flow path
 Regulatory
– Negotiation for required steps and clearance levels
– Required parallel AA testing for a period to establish safety
– Interested in product impact-immunogenicity for each strain
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Validation: Normal Operating Conditions
• MMV spike
• B Phuket and H1 California strains
• Conclusion: Clearance equivalent to previous
experimental results, complete inactivation
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Run
Run 1 H1
California
Run 2 H1
California
Log Clearance
>7.88 ± 0.07
Run 1 B Phuket
Run 2 B Phuket
>7.74 ± 0.05
>7.76± 0.07
>7.86± 0.05
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Summary
 We at Medicago have developed our strategy for containment and
control based upon Canadian, ICH and FDA guidances to the
industry
 Quality Systems and GMP principles cover the entire process
including the plant ‘biological starting material’
 The use of plants as “bioreactors” results in containment that differs
from traditional biotech operations
 Downstream operations are similar to those used in other biotech
processes and result in efficient and timely removal of bacterial
microorganisms and host cells
 Process validation and QBD principles apply to processes producing
biotherapeutics and vaccines from plants as they do to other
substrates
 While plants may be inherently safe with regard to expansion of
human pathogens, the regulatory agencies have concerns with raw
materials and vectors that may introduce adventitious agents into
plant derived products
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