Download Reference Standards for

CASSS WCBP 2012: 16th Symposium on the Interface of Regulatory and
Analytical Sciences for Biotechnology Health Products
January 23 - 25,
25 2012
Reference Standards for
Monoclonal Antibodies: Key
Challenges
g Addressed
Matthew Borer, Ph.D.
Advisor
Analytical Sciences Research and Development
Eli Lilly and Company
The Role of Reference Standards:
Pharmaceutical Analysis
Reference Standards are developed
p as p
part of the
analytical control strategy for each drug product
Each reference standard has a control strategy of
its own
Reference Standards play a central role in
assuring the quality of medicines for patients
during cGMP testing and release activities
WCBP 2012, 25-Jan-2012, M.
Borer, et al.
© 2012 Eli Lilly and Company
2
Typical RS Packaging Options
Bulk containers
dispense upon
customer order
Stable compounds
No handling issues
Less expensive materials
Infrequent
q
use
Prepackaged powder
Screw-cap bottle
Flame sealed ampo
Flame-sealed
ampoule
le
Efficient dispensing (resources &
materials)
Convenient for lab (end-use
(end use dating)
Protection from oxygen and moisture
Content per vial
Content-per-vial
Shipping “limited quantity exception”
Difficult to weigh
Hygroscopic, flocculent
Lyophilized
Expensive or limited supply
T i
Toxic
WCBP 2012, 25-Jan-2012, M.
Borer, et al.
© 2012 Eli Lilly and Company
3
Key Challenge: Packaging
Solution
So
u o p
presentation
ese a o typically
yp ca y mandatory
a da o y
Poly-cryovials
Re-closable
Re
closable
• No special filling equipment
• Compatibility testing
•
– CO2 ingress
– adsorption
– extractables
Cryo-compatible glass
ampoules
•
•
•
•
•
•
WCBP 2012, 25-Jan-2012, M.
Borer, et al.
Hermetic seal
Inert atmosphere
F
Fewer
issues
i
with
ih
adsorption/extractables
Less expensive
Single use
Special filling equipment
© 2012 Eli Lilly and Company
4
Key Challenge: Sterility
Is Sterility Required?
The reference
Th
f
standard
t d d is
i used
d iin cell-based
ll b
d assays where
h
contaminant
t i
t
organisms can proliferate
• Contaminant organisms can grow and degrade peptides on storage
•
Or Not?
Reference materials are not drug products, so patient safety is not impacted
• Aseptic manufacturing is difficult and facilities might not accept some source
materials
•
An Answer:
Sterile filter the bulk reference material source solution
• Prepare the reference materials in a clean environment
• Store reference materials frozen
• Include a broad-spectrum antibiotic in the cell culture medium
•
WCBP 2012, 25-Jan-2012, M.
Borer, et al.
© 2012 Eli Lilly and Company
5
Key Challenge:
Source Material Selection
Development
batch or batch
used in
toxicological
testing. May be
sourced
d ffrom
mixed pool of
clones.
First Laboratory
RS
Recommended to
replace the initial
batch with one
derived from the
FHD batch, or a
batch that is
representative.
New Batch
First Human Dose ((FHD))
WCBP 2012, 25-Jan-2012, M.
Borer, et al.
© 2012 Eli Lilly and Company
Replace batch only
when projected to
stock-out or in the
eventt off a significant
i ifi
t
process change.
New Batch
Commercial
Development
First Efficacy
y Dose ((FED))
6
Source Material: Commercialization
Derived from, or representative
of a batch used in pivotal clinical
of,
studies. Represents the final
optimized commercial process.
Compendial
p
RS
New Batch
Primary RS
((unlikely
y event))
Secondary RS
New Batch
Formal Stability
Prefer to source from
same packaging
k i run as
the first Primary RS.
WCBP 2012, 25-Jan-2012, M.
Borer, et al.
New Batch
Global Registration
Subsequent Secondary
RS sourced
d ffrom a
representative batch.
© 2012 Eli Lilly and Company
7
Key Challenge: Characterization
Reference standard characterization must be
customized to support its specific intended use
ICH Q3a, IV
•
Reference
R
f
standards
t d d used
d iin th
the analytical
l ti l procedures
d
ffor control
t l off iimpurities
iti should
h ld
be evaluated and characterized according to their intended uses.
uses
WHO, Annex 3, Technical Report Number 885,3
•
It is necessary to consider all data obtained from testing the material by a wide variety
of analytical methods. When taken as a whole, this will ensure that the substance is
suitable for its intended use.
use The extent of the analyses required depends on the
purpose(s) for which the chemical reference substance is to be employed, and may
involve a number of independent laboratories.
ISO 34, 4.1.1
•
It should be recognized that a reference material needs to be characterized mainly to
the level of accuracy required for its intended purpose
WCBP 2012, 25-Jan-2012, M.
Borer, et al.
© 2012 Eli Lilly and Company
8
Typical mAb Characterization Tests
Appearance
pp
Purityy
Identity
•
Light
g and Heavy
y Chain MS
Protein Quantity
•
Oligosaccharide Profile
UV
•
Charge Heterogeneity
•
Size-exclusion HPLC
• Reduced SDS-PAGE (or CESDS))
•
Capillary Electrophoresis-LIF
Potency
Ion-exchange
g chromatography
g p y
•
Bioassay
pH
WCBP 2012, 25-Jan-2012, M.
Borer, et al.
© 2012 Eli Lilly and Company
9
The Characterization Challenge
No wayy to fullyy define the Potencyy of a mAb using
g
physiochemical testing, so the Primary RS defines
biological activity
but
The Primaryy RS has no basis for comparison,
p
, so
monitoring for change in Potency is hampered
and
Bioassay methods are typically highly variable,
making it difficult to measure small changes
WCBP 2012, 25-Jan-2012, M.
Borer, et al.
© 2012 Eli Lilly and Company
10
Potency Assignment Strategy:
Initial Batch
Laboratory
RS
First Laboratoryy RS is assigned
g
a value of 100%
Relative Potency (RP)
•
Primary
RS
Subsequent Laboratory RS batches are assigned relative to the
previous Laboratory RS
The Primary RS is assigned a value of 100% RP
Confirmation of a dose response relationship
• For-information-only Bioassay comparison to Laboratory RS
•
– more extensive for IgG1 mAbs due to greater potential for potency change
•
Secondary
RS
Extensive physicochemical comparability testing to Laboratory RS
The initial Secondary RS is based on the same
characterization tests as the Primary RS (same source material)
•
A subsequent Secondary RS is assigned a value of 100% RP if the
bioassay versus the Primary RS is within a pre-defined window
WCBP 2012, 25-Jan-2012, M.
Borer, et al.
© 2012 Eli Lilly and Company
11
Potency Assignment Strategy:
Replacement Batch
Laboratory
RS
Analyzed
y
by
y the biological
g
assay
y using
g 5 independent
p
measurements relative to the previous RS
If the average relative potency is within 80-120%
80 120% RP
RP,
the new reference standard will be assigned a value of
100% RP
If the average potency falls outside of the interval, an
investigation is conducted.
Question the suitability of the material
• Consider a larger number of replicate measurements in order to
assign the RP directly to the previous standard
•
WCBP 2012, 25-Jan-2012, M.
Borer, et al.
© 2012 Eli Lilly and Company
12
Potency Assignment Strategy:
Replacement Batch
Secondary
RS
Analyzed
y
by
y the biological
g
assay
y using
g 10 independent
p
measurements relative to the Primary RS
If the average relative potency is within 95-105%
95 105% RP
RP,
the new reference standard will be assigned a value of
100% RP
If the average potency falls outside of the interval, an
investigation is conducted
Question the suitability of the material
• Consider a larger number of replicate measurements in order to
assign the RP directly to the previous standard
•
WCBP 2012, 25-Jan-2012, M.
Borer, et al.
© 2012 Eli Lilly and Company
13
Independent Measurement Defined
Independent
p
Measurement - In General
•
An average reported result based on a replication strategy that
encompasses appropriate variance components associated with the
measurement
Independent Measurement
for Bioassay
Three plates
• Three replicates at each dose
level for each:
• Sample
• Standard
• Control
C
sample
•
WCBP 2012, 25-Jan-2012, M.
Borer, et al.
© 2012 Eli Lilly and Company
14
Key Challenge: Ongoing Suitability
for Use (a.k.a.
(a k a stability)
Comprehensive physicochemical testing
Monitoring bioassay curve parameters over time
Overprotective packaging and storage temperature
Assess representative stability (e.g., from the commercial process)
Stress stability
Promote the implementation of higher order standard (e.g., WHO
International Standard)
Careful control of bioassay
•
•
•
•
Use of an independent control material
Control over key reagents
Consistent technique and comprehensive analyst training
Control over instrumentation
WCBP 2012, 25-Jan-2012, M.
Borer, et al.
© 2012 Eli Lilly and Company
15
Future Challenges
The difficulties associated with characterization of
biomolecule reference standards make harmonization of
multiple compendial reference standards a challenge
Global manufacturers must meet regional standards
• Different doses of the same drug in different regions does not make
sense for patients
• Differences in compendial RS assignments can happen
•
– Variability in measurements
– Differences in the characterization approach
This will be a particular
challenge for monoclonal
antibodies
WCBP 2012, 25-Jan-2012, M.
Borer, et al.
© 2012 Eli Lilly and Company
16
Co Authors
Co-Authors
Kimberley
y B. Dancheck
Kristi L. Griffiths
Bryan J. Harmon
Jerry J. Lewis
Jerry Zhirui Lian
Bhavin S. Parekh
WCBP 2012, 25-Jan-2012, M.
Borer, et al.
© 2012 Eli Lilly and Company
17