Analytical Control Strategy for robust vaccine …...Strategy for robust vaccine development...
Transcript of Analytical Control Strategy for robust vaccine …...Strategy for robust vaccine development...
Analytical Control Strategy for robust vaccine development
Cristiana CampaWCBP, January 2016
Key elements for a robust Analytical Control StrategyBuild strong analytical knowledge beginning with early development, leading to a systematic increase of product and process understanding during life cycle
Refine
Testing
Strategy
during life cycle
Execute method development/ performance
verification
Screen and select analytical technology
Ensure compliance of analytical methods with current product and process requirements
(Analytical Target Profile)
2
Management of product & process expectation through Analytical Target Profile
• Well- defined (possibly fixed) methods are desired since early development to ensure visibility of changes induced during product & process development/ life cycle
Product & Process«clients» expectation
• Minimal change of methods occur during all life cycle, with some resistance to changes or improvementsConsequences
• Product and process requirements are built over time (platform concept is not fully in place for vaccines)
• Always updated analytical technologies are neededgiven vaccines complexity
Challenges for vaccines
• Analytical Target Profile (ATP) is defined with clients since early development, including ideal performances of the analytical methods based on current product and process requirements
Solution
3
Input for ATP- product understanding
Serotype III
Risk assessment for identification of CQAs is important to ensure prioritization of analytical activities
Sialic acid residue is susceptible to hydrolysis; hydrolysis rate depends on temperature and conditions (e.g. pH) sialic acid cleavage changes immunological epitope
Case study Group B Streptococcus glycoconjugate
Focus analytical effort on relevant aspectsExample: for product analytics the input for ATP is Quality Target Product Profile
(Critical Quality Attributes (CQAs) and ranges as applicable)
4
Section 1. General Information
Sample Intended Purposes of Measurement Scope Category Output
Drug Product
Measure free sialic acid as CQA and
stability indicator to be monitored in
stability studies
1. Measure a CQA
2. Stability
Quantitative test for a product
related impurity
Section 2. Performance Requirements (related to the most challenging purpose)
SpecificityAccuracy
(meaning of)Accuracy Rationale Precision Rationale
Able to
discriminate
analyte from
matrix signals
Concordance with
the true value80-120%
Adequate to current
knowledge about
effect of sialic acid
cleavage on
immunogenicity
CV≤10%
Adequate for a
stability indicator
method considering
current specs limit
A practical example of specifications settingExtract from ATP
Case study: Group B Streptococcus glycoconjugate- free sialic acid in DP
Note: Business considerations (eg throughput, cycle time, ...) are also included in ATP, but they are not shown here
5
Example of analytical technology selection
• 1H-NMR: sialicacid retention in release
Polysaccharide
• Sialic acid not in release specification panel but is a characterization assay; it has been demonstrated that process steps do not alter sialic acid retention attribute
Drug Substance
• HPAEC-PAD: free sialic acid at release and in stability
• Potency assay sensitive to sialic acid loss
Drug Product
Sialic acid
Standard)
Sample
HPAEC-PAD
V. Pinto; F. Berti;
Biomed. Anal. 98
(2014);9-15
Case study: Group B Streptococcus glycoconjugate- free sialic acid
Carefully define methods application/ purposebased on product characteristics and ability to control CQAs from process
6
Case study: characterization of Rotavirus
Thorough characterization can be used for methods selection but also to confirm structural features to be monitored -this may need more than one method
•Low resolution of DLS makes it not applicable for real samples with mixed DLP/TLP •EM can discriminate DLP and TLP, and also detect damaged/empty particles, but high number of particles should be observed and sized to get accurate relative abundances •DC and CE are medium throughput techniques suitable for quantitative assay, and able to baseline resolve DLP from TLP
Triple-layered particles containing all 3 protein layers (TLP), as well as smaller, double-layered particles lacking outer capsid proteins VP4 and VP7 (DLP), are detected in preparations of live attenuated rotavirus for vaccination.
Example of analytical technology selection
DLP are non-infectious and it may therefore be of importance to address their abundance vs. TLP in vaccine preparations.
White arrow: full particle; black arrow: damagedparticle; white-dot arrow: whole empty particle.
Negative Staining Electron Microscopyof Rotavirus
DLP TLP
Diameter of Rotavirus double- and triple-layered particles measured by Electron Microscopy, Dynamic Light Scattering, Disc Centrifuge, Capillary Electrophoresis.
Analysis of Rotavirus DLP and TLP by Disc Centrifuge (A) and Capillary Electrophoresis (B)
7
Analytical methods used for process understanding may have different requirementswith respect to those used for release/ stability testing,
although they can be applied to the same CQAs
For instance, process «screening» assesses the Process Parameters criticality (i.e., variation of CQA(s) upon changing PP value), not the evaluation of the impact of the change on product quality within specifications (which is the aim of «mapping» studies)
Therefore the typical desired features for methods used for screening studies are- High throughput (to support multivariate studies and to efficiently monitor process)- High precision (to ensure reliability of assessed CQA changes due to PP changes)- High selectivity for complex matrices (to execute testing as close as possible to the investigated step)
ATP for a CQA can incorporate different applications (release/ staibility or process testing),and can potentially be associated to more than one method, if the release/ stabilityapproach is not fully suited for all applications (worse case scenario)
Process Understanding and the ATP
8
Method parameters (ATP
requirements for
performance)
Colorimetric sialic acid HPAEC-PAD with CarboPac PA1 HPAEC-PAD with CarboPac PA20Fast
Obtained value ATP requirement Obtained value ATP requirement Obtained valueATP
requirement
Selectivity / Specificity
(Able to discriminate
analyte from matrix
signals)
Analyte not separated
from other
components;
ultrafiltration needed
for complex matrix
phases
Met upon sample UF
Analyte peak
separated from the
other matrix
components
Fully met, no UF
Analyte peak
separated from the
other matrix
components
Fully met, no UF
Accuracy
(80-120%)
82-100%
(pre-validation data)Met
97-108% (pre-
validation data)Met
95-99%
(validation data)Met
Precision
(CV < 8%)
2-10%
Possible variability
between different lots
and analyses performed
in long different time
(pre-validation data)
Not fully metCV 3-10%
(pre-validation data)Not fully met
CV 3%
(validation data)Met
Cycle Time
(< 1 day )
2 hours (no
ultrafiltration)5 hours
(ultrafiltration required)
Met 10-12 hours Met 4-5 hours Met
Throughput
(NLT 8 samples)Up to 20 Met Up to 15 Met Up to 30 Met
Sample volume required
for the analysis1 ml - 20 µl - 5 µl -
Product development stages
Case study: Group B Streptococcus glycoconjugate- saccharide content
F. Merangolo, S. Giannini, M. Gavini, S. Ricci, C. Campa, LCGC Applications of Ion Chromatography (2015) 9
Process Understanding and the ATP
Key elements for a robust Analytical Control StrategyBuild strong analytical knowledge beginning with early development, leading to a systematic increase of product and process understanding during life cycle
Refine
Testing
Strategy
during life cycle
Execute method development/ performance
verification
Screen and select analytical technology
Ensure compliance of analytical methods with current product and process requirements
(Analytical Target Profile)
10
After analytical technology selection, risk of failure can be limited through:
Method development including QbD elements (eg applying a risk- basedapproach and, possibly, multi-variate studies), with final verification of performances, to be compliant with ATP
Establishment of a robust reference standard strategy (thoroughcharacterization, stability studies/ definition of storage conditions, use for method performance monitoring)
Note: these activities are critical also to ensure adequate bridging betweenmethods, if needed during life cycle management (eg for introduction of new technologies, or if ATP is refined after QTPP revision duringdevelopment)
Method development and reliability assessment
11
Method development including QbD elements
S. Orlandini, S. Pinzauti, S. Furlanetto: Anal. Bioanal. Chem. 405 (2013) 443-450
Analytical Target Profile &
method selection
Quality Risk Assessment
Knowledge SpaceInvestigation by DoE
Design SpaceResponse Surface Metodology
Method Control
Working PointsRobustness
12
Case study: Quality Risk Assessment (QRA) - UPLC for % adsorption of protein on an aluminum surface
Ishikawa diagram (fishbone) for screening the method parameters andfor identifying the critical process parameters (CPP) to be furtherstudied by DoE.
Screening study of the effects of chromatographic parameters onchromatographic performances
Method development including QbD elements
13
287-
953
Por
B
Por
A
936-
741
961c
AU
0,00
0,10
0,20
0,30
0,40
0,50
0,60
0,70
0,80
0,90
1,00
1,10
1,20
1,30
Minutes
3,00 3,10 3,20 3,30 3,40 3,50 3,60 3,70 3,80 3,90 4,00 4,10 4,20 4,30 4,40 4,50 4,60 4,70 4,80 4,90 5,00 5,10 5,20 5,30 5,40 5,50 5,60 5,70 5,80 5,90 6,00
Pro
tein
I
Pro
tein
II
Pro
tein
III
Pro
tein
V
Pro
tein
IV
Case study: knowledge space- UPLC for % adsorption of protein on an aluminum surface
Method development including QbD elements
Asymmetric matrix (based onFree-Wilson model) forinvestigation of the KnowledgeSpace (KS) and to obtainpreliminary information on theeffects of the factors on methodperformance (eg peakresolution and area).
Re
solu
tio
nP
rote
in I-
Pro
tein
II
14
Case study: knowledge space - UPLC for % adsorption of protein on an aluminum surface
Method development including QbD elements
Investigated performance indicators
R1: Protein I – Protein II Resolution
R2: Protein II– Protein III Resolution
R3: Protein III – Protein IV Resolution
R4: Protein V – isoform Resolution
Study outcome:
Parameters impacting peak resolution and areas & best conditions found at this stage:
- Column type: C4 pore best column
- % start Acetonitrile: 34% best value based on screening studies (under verification -design space assessment)
- Ramp time: 4 or 6 min best values based on screening studies (under verification -design space assessment)
- Column temperature: 60°C best value based on screening studies (under verification –design space assessment)
A1: Protein II
A2: Protein III
A3: Protein IV
15
Key elements for a robust Analytical Control StrategyBuild strong analytical knowledge beginning with early development, leading to a systematic increase of product and process understanding during life cycle
Refine
Testing
Strategy
during life cycle
Execute method development/ performance
verification
Screen and select analytical technology
Ensure compliance of analytical methods with current product and process requirements
(Analytical Target Profile)
16
Methods life cycle
Analytical testing strategy evolves with process and product knowledge,
and doesn’t stop with Regulatory Approval.
Post-Licensure changes in the analytics:
may be driven by several factors focused to maintain the best control on the product and process
must consider pros and cons for total impact on strategy
Pro
s • Performance improvement (robustness, cycle time, throughput, cost)
• Innovation advancing• Harmonization between
labs/sites• Ethical Change from in vivo to in
vitro• Deepening product Knowledge
Ch
alle
nges • Equivalence demonstration and
Impact on specification
• Regulatory variations required
• Alignment with OMCLs
• Internal resistances
• Bridging with historical data
• Use for stability studies
17
– A test was required to assess carbohydrate content in polysaccharides.
– The Resorcinol assay is based on the acidic dehydration of monosaccharides (hexoses) to 5-HMF.
– 5-HMF reacts with Resorcinol to generate a chromophore whose concentration is determined using a UV-visible spectrometer (usually 430nm).
– In its apparent simplicity, this assay hides several drawbacks.
– The generation of 5-HMF is continuous until exhaustion of the carbohydrate => the signal evolves.
– The kinetic of a monosaccharide is not necessarily the same than a polysaccharide.
– It will have consequences on the accuracy and on the standard choice
– There are other methods allowing to achieve more information with respect to colorimetric approach
5-HMF
Resorcinol
Hexoses
Chromophore
NMR vs colorimetric – Pneumococcal polysaccharide 23F
Methods life cycle
18
Easy sample preparation
Accurate
Specific
Precise
All in one
Easy Standard management
Inte
rna
l S
tan
da
rd
CPS
Acetate
Methyl
Pentose
EtOH
Silicon
Anomeric
RegionH
DO
Identity
Aromatic Amino Acid
R/DNA Base
Phenol
NMR vs colorimetricPro: innovation 1H Nuclear Magnetic ResonanceNMR vs colorimetric – Pneumococcal polysaccharide 23F
Methods life cycle
19
Analytical Control Strategy & Life Cycle
Analytical Target Profile
Analytical Methods Screening/ Development/ Refinement (for release/ stability & characterization)
Ph IIIPh IIPh IDiscovery Preclinical Registr.Launch
Reference standard (selection, characterization and stability)
Analytical methods qualification/ validation
Criticality confirmation studies for quality attributes and Physico- chemical comparability
Analytical Methods Screening/ Development/ Refinement (for process understanding)
Refinement
QTPP & process
requirements
Analytical Methods Life cyclemanagement/ development continuity
Refinement
Characterization
efforts since
early
development
Final ATP Submission
20
Challenges & Opportunities
For vaccines, product/ process requirements are defined/ refined over time with consequent need to change ATP and, potentially, selected methods
o Optimization is ensured through screening of orthogonal methods when product / process requirements are under assessment (this is also needed to minimize risks of undetected issues/ identify CQAs)
o Extensive characterization efforts are needed since early development. Apparent investment of time/ resources is compensated by robustunderstanding and control of manufacturing aimed at ensuring product quality.
After fixing product / process requirements (eg typically in late development/ commercial phase), with a final/fixed ATP, it is important to update analytical technologies, to ensure the best testing approach for vaccines at any stage of life cycle.
o Regulatory filing could be effected upon possibility to submit Analytical Target Profile.
21
Acknowledgement
Amin KhanBill EganDominique LemoineDaniela StrangesGhislain DelpierreThomas JacquesLuca NompariAuthors of the mentioned publications
22