outcomes of the global bioanalysis consortium s ... · outcomes of the global bioanalysis...
TRANSCRIPT
![Page 1: Outcomes of the Global Bioanalysis Consortium s ... · Outcomes of the Global Bioanalysis Consortium’s Recommendations: Large Molecule and All Molecules ... Recommended a draft](https://reader033.vdocument.in/reader033/viewer/2022060223/5f07cad67e708231d41ec4e9/html5/thumbnails/1.jpg)
Outcomes of the Global Bioanalysis Consortium’s Recommendations: Large
Molecule and All Molecules Harmonization teams
Webinar - May 2013
![Page 2: Outcomes of the Global Bioanalysis Consortium s ... · Outcomes of the Global Bioanalysis Consortium’s Recommendations: Large Molecule and All Molecules ... Recommended a draft](https://reader033.vdocument.in/reader033/viewer/2022060223/5f07cad67e708231d41ec4e9/html5/thumbnails/2.jpg)
A team
2
![Page 3: Outcomes of the Global Bioanalysis Consortium s ... · Outcomes of the Global Bioanalysis Consortium’s Recommendations: Large Molecule and All Molecules ... Recommended a draft](https://reader033.vdocument.in/reader033/viewer/2022060223/5f07cad67e708231d41ec4e9/html5/thumbnails/3.jpg)
A1 – Scope and Regulations
Defined and contrasted Regulations, Guidance and White
papers
Regardless of GxP, recommendations made for performing
Regulated BA with an hierarchical structure – Validations to follow regulated BA principles
– Samples analysis following regulated BA principles, while maintaining
GLP and GCP status for samples
Recommended a draft scope statement for GBC
whitepaper – Clarified scope for regulated bioanalysis
– Included timelines for validations and scope (extent) of validation before
analysis of samples
Created draft glossary and abbreviations for BA
3
![Page 4: Outcomes of the Global Bioanalysis Consortium s ... · Outcomes of the Global Bioanalysis Consortium’s Recommendations: Large Molecule and All Molecules ... Recommended a draft](https://reader033.vdocument.in/reader033/viewer/2022060223/5f07cad67e708231d41ec4e9/html5/thumbnails/4.jpg)
Discovery
IND GLP tox
Ph1
Ph 3
Post approval
Ph0
Ph 1 or 2 & Chronic GLP tox
Non-GLP Preclinical or in vitro
Ph 2
Screening, research or qualified methods as desired
Parent drug, prodrug or metabolite in any matrix: qualified method method
Parent drug in plasma: validated method
Parent drug in other matrices: qualified method
Metabolite/Prodrug in any matrix: qualified method
Human vs. animal plasma for MIST: screening method
method
Parent drug & post-MIST* metabolite in plasma: validated method
Parent drug & post-MIST metabolite in other matrices: qualified method
A2 – Tiered Approach for MV
4
![Page 5: Outcomes of the Global Bioanalysis Consortium s ... · Outcomes of the Global Bioanalysis Consortium’s Recommendations: Large Molecule and All Molecules ... Recommended a draft](https://reader033.vdocument.in/reader033/viewer/2022060223/5f07cad67e708231d41ec4e9/html5/thumbnails/5.jpg)
A2 – Tiered Approach for MV
Practicality in 483 averse environment
4 tiers proposed (screening, research,
qualified, validated)
When is each tier applied?
5
![Page 6: Outcomes of the Global Bioanalysis Consortium s ... · Outcomes of the Global Bioanalysis Consortium’s Recommendations: Large Molecule and All Molecules ... Recommended a draft](https://reader033.vdocument.in/reader033/viewer/2022060223/5f07cad67e708231d41ec4e9/html5/thumbnails/6.jpg)
A3 – Method Transfer and Cross Validation
•Transfer is a process not a cross validation
– Internal: Two P/A batches (4 for LBA)
– External: Full validation & inter-lab comparison (“may” included spiked QC’s)
•Matrix stability not repeated in cross/partial validations
•Cross validation consists of analysis of assessment samples (spiked QCs and incurred samples) assayed using two or more different validated methods
6
![Page 7: Outcomes of the Global Bioanalysis Consortium s ... · Outcomes of the Global Bioanalysis Consortium’s Recommendations: Large Molecule and All Molecules ... Recommended a draft](https://reader033.vdocument.in/reader033/viewer/2022060223/5f07cad67e708231d41ec4e9/html5/thumbnails/7.jpg)
A4 – Reference Standards and Reagents
Calibrators and QC samples should be
prepared from a verified stock solution with
proven stability
Altered or surrogate matrix should only be
used if equivalency is demonstrated
between surrogate matrix and authentic
matrix. QC samples should be prepared in
authentic matrix
7
![Page 8: Outcomes of the Global Bioanalysis Consortium s ... · Outcomes of the Global Bioanalysis Consortium’s Recommendations: Large Molecule and All Molecules ... Recommended a draft](https://reader033.vdocument.in/reader033/viewer/2022060223/5f07cad67e708231d41ec4e9/html5/thumbnails/8.jpg)
A4 – Reference Standards and Reagents
New reference standard qualification
• Qualified via single partial validation run with L,
M, H validation QC samples (n=6)
Alternate: Stds and QC’s from each lot with
QC’s read from each curve.
8
![Page 9: Outcomes of the Global Bioanalysis Consortium s ... · Outcomes of the Global Bioanalysis Consortium’s Recommendations: Large Molecule and All Molecules ... Recommended a draft](https://reader033.vdocument.in/reader033/viewer/2022060223/5f07cad67e708231d41ec4e9/html5/thumbnails/9.jpg)
• Sample collection at testing site
• Storage and shipment from testing site to analytical
lab
• Receipt of samples by receiving lab
• Storage at analytical lab (pre and post analysis)
• Temperature ranges
• Sample management using LIMS
• Sample disposal
A5: Sample Management - Topics discussed
![Page 10: Outcomes of the Global Bioanalysis Consortium s ... · Outcomes of the Global Bioanalysis Consortium’s Recommendations: Large Molecule and All Molecules ... Recommended a draft](https://reader033.vdocument.in/reader033/viewer/2022060223/5f07cad67e708231d41ec4e9/html5/thumbnails/10.jpg)
A5: Sample Management - Summary
10
• An audit trail of the location and conditions of the
samples must be maintained from collection to
disposal
• Storage conditions must be monitored at all times :-
Ambient, Fridge, Freezer, Ultracold
(The group has recommendations for the temperature
ranges, furthermore the use of the terms -70 or -80
storage should not be used)
• Temperature excursions outside the acceptance
ranges must be documented together with any impact
• Clinical samples must be disposed of at the end of the
trial
![Page 11: Outcomes of the Global Bioanalysis Consortium s ... · Outcomes of the Global Bioanalysis Consortium’s Recommendations: Large Molecule and All Molecules ... Recommended a draft](https://reader033.vdocument.in/reader033/viewer/2022060223/5f07cad67e708231d41ec4e9/html5/thumbnails/11.jpg)
A6 – Stability
Stability in WB not required unless
scientifically called for ; assumes
plasma/serum stability under the same
conditions
Stability in the presence of co-medications
or in unique matrix (e.g. hemolytic) not
required unless scientifically called for.
11
![Page 12: Outcomes of the Global Bioanalysis Consortium s ... · Outcomes of the Global Bioanalysis Consortium’s Recommendations: Large Molecule and All Molecules ... Recommended a draft](https://reader033.vdocument.in/reader033/viewer/2022060223/5f07cad67e708231d41ec4e9/html5/thumbnails/12.jpg)
A6 – Stability
Use of freshly prepared calibrators
recommended for long-term stability only
Incurred sample stability not required as a
standard. It should serve to bridge a
possible gap between spiked and incurred
samples, when deemed necessary based
on the physicochemical and /or metabolic
properties of the analyte.
12
![Page 13: Outcomes of the Global Bioanalysis Consortium s ... · Outcomes of the Global Bioanalysis Consortium’s Recommendations: Large Molecule and All Molecules ... Recommended a draft](https://reader033.vdocument.in/reader033/viewer/2022060223/5f07cad67e708231d41ec4e9/html5/thumbnails/13.jpg)
A7 – Reanalysis and ISR
Incongruous repeats
• Select prior to PK analysis
• Repeat in duplicate
• Accept if reassay if difference < 30% ;
< 40% for LBA
(Feedback needed, align with ISR %?)
• Report median of 3 results
13
![Page 14: Outcomes of the Global Bioanalysis Consortium s ... · Outcomes of the Global Bioanalysis Consortium’s Recommendations: Large Molecule and All Molecules ... Recommended a draft](https://reader033.vdocument.in/reader033/viewer/2022060223/5f07cad67e708231d41ec4e9/html5/thumbnails/14.jpg)
A7 – Reanalysis and ISR
ISR
• Largely a QC effort, scientific return obtained
after first test per unique matrix
Amount of ISR
• 5% of samples for all studies (eliminate tiers)
• Minimum of 20 samples
ISR Failures
• How to deal with individual outliers
14
![Page 15: Outcomes of the Global Bioanalysis Consortium s ... · Outcomes of the Global Bioanalysis Consortium’s Recommendations: Large Molecule and All Molecules ... Recommended a draft](https://reader033.vdocument.in/reader033/viewer/2022060223/5f07cad67e708231d41ec4e9/html5/thumbnails/15.jpg)
A7 – Reanalysis and ISR
ISR Selection
• Based on visual inspection, low/high
concentration range of samples, > 3XLLOQ
ISR for Multi-analyte methods
• Select based on primary active entity
• Consideration to other analytes, but not driven
by them unless they represent a major
metabolite or co-med and are largely separated
pharmacokinetically
15
![Page 16: Outcomes of the Global Bioanalysis Consortium s ... · Outcomes of the Global Bioanalysis Consortium’s Recommendations: Large Molecule and All Molecules ... Recommended a draft](https://reader033.vdocument.in/reader033/viewer/2022060223/5f07cad67e708231d41ec4e9/html5/thumbnails/16.jpg)
A8 – Documentation
High level table of contents proposed for the
analytical and validation report
Recommendation to reserve a separate
section in CTD on bioanalysis to avoid
confusion and help reviewers locating the
data.
16
![Page 17: Outcomes of the Global Bioanalysis Consortium s ... · Outcomes of the Global Bioanalysis Consortium’s Recommendations: Large Molecule and All Molecules ... Recommended a draft](https://reader033.vdocument.in/reader033/viewer/2022060223/5f07cad67e708231d41ec4e9/html5/thumbnails/17.jpg)
Discussion Topics
• Dilution QC’s
• Reinjection reprod./stability
• IS reproducibility
• Integration
• Selectivity/carryover
% LLOQ vs. LLOQ perf.
• System equilibration,
suitability, requalif.
• Tiered MV
• Method transfer/cross val.
• Ref std prep and qualif.
• Stability: ISS, WB, co-meds
17
• Repeat analysis for incongruous
results
• ISR amount, selection and
individual sample failures
![Page 18: Outcomes of the Global Bioanalysis Consortium s ... · Outcomes of the Global Bioanalysis Consortium’s Recommendations: Large Molecule and All Molecules ... Recommended a draft](https://reader033.vdocument.in/reader033/viewer/2022060223/5f07cad67e708231d41ec4e9/html5/thumbnails/18.jpg)
Not part of this webinar
18
A9 – Instrument Qualification
![Page 19: Outcomes of the Global Bioanalysis Consortium s ... · Outcomes of the Global Bioanalysis Consortium’s Recommendations: Large Molecule and All Molecules ... Recommended a draft](https://reader033.vdocument.in/reader033/viewer/2022060223/5f07cad67e708231d41ec4e9/html5/thumbnails/19.jpg)
A10 – New Frontiers
Not part of this webinar
19
![Page 20: Outcomes of the Global Bioanalysis Consortium s ... · Outcomes of the Global Bioanalysis Consortium’s Recommendations: Large Molecule and All Molecules ... Recommended a draft](https://reader033.vdocument.in/reader033/viewer/2022060223/5f07cad67e708231d41ec4e9/html5/thumbnails/20.jpg)
A11 – Biomarkers
Not part of this webinar
20
![Page 21: Outcomes of the Global Bioanalysis Consortium s ... · Outcomes of the Global Bioanalysis Consortium’s Recommendations: Large Molecule and All Molecules ... Recommended a draft](https://reader033.vdocument.in/reader033/viewer/2022060223/5f07cad67e708231d41ec4e9/html5/thumbnails/21.jpg)
L teams
21
![Page 22: Outcomes of the Global Bioanalysis Consortium s ... · Outcomes of the Global Bioanalysis Consortium’s Recommendations: Large Molecule and All Molecules ... Recommended a draft](https://reader033.vdocument.in/reader033/viewer/2022060223/5f07cad67e708231d41ec4e9/html5/thumbnails/22.jpg)
L1 - LBA Run Acceptance
The team’s recommendations on acceptance criteria employed both in
validation of ligand binding methods and during sample analysis are well
aligned with relevant published regulatory guidance and white papers
Consensus achieved on:
Use of Total Error during pre-study validation to define in-study QC
acceptance criteria
Prepare, qualify and freeze aliquots of standard curve calibrators.
Establish stability by comparing fresh vs. frozen standard curve
calibrators during A&P.
When masking a calibrator is employed, it is strongly
recommended that an objective, step-by-step process be defined a
priori
Quality Controls should be prepared separately from calibrators,
qualified and frozen to mimic the study samples. QCs should
always be used for validation and analysis from the frozen state.
![Page 23: Outcomes of the Global Bioanalysis Consortium s ... · Outcomes of the Global Bioanalysis Consortium’s Recommendations: Large Molecule and All Molecules ... Recommended a draft](https://reader033.vdocument.in/reader033/viewer/2022060223/5f07cad67e708231d41ec4e9/html5/thumbnails/23.jpg)
Recommendations:
•Ensure same number of observations in each Precision & Accuracy (P&A) run – number of sets of QCs in each run should be the same
• Aligned with Team L1, which recommends 3 independent sets each of 5 QC levels (LLOQ, LQC, MQC, HQC, ULOQ) in ≥ 6 P&A runs
•Analyst as variable: Number of analysts in validation should be reflective of sample testing practice, noting that:
• Commonly one analyst performs bulk of validation with inclusion of second analyst for a subset of P&A runs
• Possible to justify using only one analyst during validation if study sample analysis will also only use one analyst (small studies)
• If multiple analysts will test samples, recommend at least 2 analysts in validation
L2 - Setting Up Balanced Validation
Design
![Page 24: Outcomes of the Global Bioanalysis Consortium s ... · Outcomes of the Global Bioanalysis Consortium’s Recommendations: Large Molecule and All Molecules ... Recommended a draft](https://reader033.vdocument.in/reader033/viewer/2022060223/5f07cad67e708231d41ec4e9/html5/thumbnails/24.jpg)
L2 - Dilution Linearity and Hook Effect
Recommendations:
Approach •Spike samples at ≥ anticipated Cmax; if Cmax is unknown, spike at highest feasible concentration •There should be a minimum of 90% matrix in highest feasible concentration sample •Make dilution series that includes anticipated sample concentrations both above and spanning the curve range
Acceptance Criteria •In-range samples should be within 20% theoretical and precision of the cumulative back-calculated concentrations should be ≤ 20% •If >ULOQ spiked samples read ALQ = no hook effect
![Page 25: Outcomes of the Global Bioanalysis Consortium s ... · Outcomes of the Global Bioanalysis Consortium’s Recommendations: Large Molecule and All Molecules ... Recommended a draft](https://reader033.vdocument.in/reader033/viewer/2022060223/5f07cad67e708231d41ec4e9/html5/thumbnails/25.jpg)
Risk based approach
• A case by case risk assessment is needed to determine whether in-study hook effect evaluation should be done
Recommendation:
• The scientist should be mindful of any special characteristics or handling procedures related to the drug and how this might lead to potential hook effect in study samples even when one is not observed during validation
• In study, the scientist should be reviewing the data closely in order to “catch” any hook effect (as well as loss of parallelism) that may manifest
L2 - In-study Hook Effect
![Page 26: Outcomes of the Global Bioanalysis Consortium s ... · Outcomes of the Global Bioanalysis Consortium’s Recommendations: Large Molecule and All Molecules ... Recommended a draft](https://reader033.vdocument.in/reader033/viewer/2022060223/5f07cad67e708231d41ec4e9/html5/thumbnails/26.jpg)
L2 - Testing Robustness and Ruggedness Definition • Robustness/ruggedness terms are often used interchangeably and there has
historically been confusion regarding the absolute definition of each term • It is understood, however, that both parameters are indicators of assay
reproducibility under varied conditions • Therefore, any robustness/ruggedness analysis should address the question
of whether the assay will perform well under real life changes in standard laboratory situations
Recommendations • Robustness/ruggedness testing should generally be incorporated into
method development process • Typically includes temperature variations, reagent lots, plate lots, analysts,
instruments, incubation times • Be aware of the needs of your assay and the conditions under which it may be
run, including differences in ambient temperatures in different labs; regional differences in serum/plasma sources, etc.
• Demonstrated during validation by virtue of use of multiple instruments/analysts and typical variations in incubation times
• Cross validation in later stage further demonstrates R&R
![Page 27: Outcomes of the Global Bioanalysis Consortium s ... · Outcomes of the Global Bioanalysis Consortium’s Recommendations: Large Molecule and All Molecules ... Recommended a draft](https://reader033.vdocument.in/reader033/viewer/2022060223/5f07cad67e708231d41ec4e9/html5/thumbnails/27.jpg)
L2 - Selectivity
Recommendations
Approach • Test 10 or more individual samples, unspiked and spiked at LLOQ
level (required) and higher level (e.g. HQC) (recommended)
• When possible, for disease indications, selectivity should be performed in disease matrix
Acceptance Criteria • ≥ 80% of unspiked samples should measure <LLOQ
• ≥ 80% (8/10) samples should be within 20% nominal for HQC spikes and 25% for LLOQ spikes
• The same 80% of samples should meet criteria
![Page 28: Outcomes of the Global Bioanalysis Consortium s ... · Outcomes of the Global Bioanalysis Consortium’s Recommendations: Large Molecule and All Molecules ... Recommended a draft](https://reader033.vdocument.in/reader033/viewer/2022060223/5f07cad67e708231d41ec4e9/html5/thumbnails/28.jpg)
L2 - Selectivity Lipemic Samples
• Recommendation – risk based approach • The need to perform selectivity assessments with lipemic samples will
be dependent upon the drug, disease indication and assay format
• Typically, performing selectivity assessments with disease matrix samples will address any effects of lipemia which may be present in that population
Hemolyzed Samples – risk based approach
• Recommendation
• The need to perform selectivity assessments with hemolyzed samples will
be dependent upon the characteristics of drug, its target, disease
indication and assay format
• The team actively sought examples where there was an issue caused by
hemolysis to guide when these assessments may be recommended
• Examples gathered to date indicate that issues due to hemolysis are rare and
have not been observed with mAb therapeutics. However, insulin and related
therapeutics may be more likely to be sensitive to hemolysis
![Page 29: Outcomes of the Global Bioanalysis Consortium s ... · Outcomes of the Global Bioanalysis Consortium’s Recommendations: Large Molecule and All Molecules ... Recommended a draft](https://reader033.vdocument.in/reader033/viewer/2022060223/5f07cad67e708231d41ec4e9/html5/thumbnails/29.jpg)
L2 - Selectivity – Other Issues
• When endogenous analyte is present
• Choosing standard curve matrix - when possible, screen for low
endogenous level pool
• Employing a subtraction method to evaluate spike recovery may be possible if endogenous levels are measureable (≥ LLOQ)
• May need to sacrifice LLOQ level when endogenous levels are detectable, but not quantifiable (< targeted LLOQ)
• If endogenous level is quite high, then may need to question if your assay range is appropriate
![Page 30: Outcomes of the Global Bioanalysis Consortium s ... · Outcomes of the Global Bioanalysis Consortium’s Recommendations: Large Molecule and All Molecules ... Recommended a draft](https://reader033.vdocument.in/reader033/viewer/2022060223/5f07cad67e708231d41ec4e9/html5/thumbnails/30.jpg)
• Routine parallelism assessments currently not being broadly implemented industry-wide (some/few are doing routinely)
• More questions than answers remain around when to perform the assessment, how to perform it, and how to report the data
• Recommendation – risk based approach • The need to perform parallelism assessments will depend upon the
characteristics of the drug, its binding partners and specific assay reagents
• The team sought examples where non parallelism had been observed to guide when assessment may be recommended
• Examples indicate that non-mAb therapeutics, especially peptides, may be more likely to have issues of non parallelism
• No examples yet identified for mAb therapeutics
L2 - Parallelism
![Page 31: Outcomes of the Global Bioanalysis Consortium s ... · Outcomes of the Global Bioanalysis Consortium’s Recommendations: Large Molecule and All Molecules ... Recommended a draft](https://reader033.vdocument.in/reader033/viewer/2022060223/5f07cad67e708231d41ec4e9/html5/thumbnails/31.jpg)
L2 - Specificity Testing
Recommendations Approach
• What to test
• Potentially cross-reacting molecule(s); Note: more relevant for non mAb drugs than mAb drugs as selectivity assessments already assess recovery in presence of mg/mL levels of antibodies
• Concomitant medications: As appropriate – test those that are specified in the protocol to be co-administered or stable regimen; No need to test OTC drugs
• Other: Consider potential impact of ADAs, circulating soluble target, etc. on assay performance
• How to test
• Spike maximum concentration anticipated in study into (1) blank matrix, (2) LLOQ QC; (3) HQC
Acceptance Criteria
• Unspiked samples should measure <LLOQ
• Spiked samples should measure within 20% nominal for HQC spikes and 25% for LLOQ spikes
![Page 32: Outcomes of the Global Bioanalysis Consortium s ... · Outcomes of the Global Bioanalysis Consortium’s Recommendations: Large Molecule and All Molecules ... Recommended a draft](https://reader033.vdocument.in/reader033/viewer/2022060223/5f07cad67e708231d41ec4e9/html5/thumbnails/32.jpg)
L3: Non-plate based Assay Platforms
Sample testing is run in series (ie Gyrolab, RIA, Biacore) on some platforms
rather than plates
Recommendation:
A run is not limited to ’96’ in this case.
short-term stability data will be used to determine the time required for
‘new runs’ to be started and drift effects if applicable
intermittent QC sets to be added during the sample analysis
A standard curve can be added in at the start of every run.
Carry over should be avoided and the method adjusted accordingly
Singlet analysis of samples can be explored as long as the CV of QC
sets is within an acceptable range developed during assay validation.
As this is the most controversial recommendation, it will require sharing
of experience with validation of larger numbers of QC sets before being
implemented widely.
![Page 33: Outcomes of the Global Bioanalysis Consortium s ... · Outcomes of the Global Bioanalysis Consortium’s Recommendations: Large Molecule and All Molecules ... Recommended a draft](https://reader033.vdocument.in/reader033/viewer/2022060223/5f07cad67e708231d41ec4e9/html5/thumbnails/33.jpg)
L3: Assay Platforms
Both transfer between different platforms and multiplexing was also discussed and
surveyed on.
Recommendation:
Method transfer: used of spiked samples to cross-validate platforms was deemed
the most reliable way of method transfer.
Multiplexing best practice: It is recommended that analytes be validated in a
cocktail whenever possible. During sample analysis, if one analyte fails then all
samples should be rerun and mask the previous passing analyte results.
![Page 34: Outcomes of the Global Bioanalysis Consortium s ... · Outcomes of the Global Bioanalysis Consortium’s Recommendations: Large Molecule and All Molecules ... Recommended a draft](https://reader033.vdocument.in/reader033/viewer/2022060223/5f07cad67e708231d41ec4e9/html5/thumbnails/34.jpg)
L3: Cell-based assays for PK
More extensive method characterization is required due to the sensitivity of the
assay to numerous factors:
Recommendation for assay characterization:
serum lots, cell passages, plating density, the length of time in culture
conditions to be monitored for assay robustness
sample should be run in triplicate
sample analysis requires that QCs be placed within the plate as well as
along the edge so that any border effects can be determined.
LLOQ, LQC, MQC, HQC, and ULOQ precision and accuracy criteria up to
30% and total error up to 40% may be required.
![Page 35: Outcomes of the Global Bioanalysis Consortium s ... · Outcomes of the Global Bioanalysis Consortium’s Recommendations: Large Molecule and All Molecules ... Recommended a draft](https://reader033.vdocument.in/reader033/viewer/2022060223/5f07cad67e708231d41ec4e9/html5/thumbnails/35.jpg)
L3: Cell based Assays for PK
Recommendations for cell line stability:
Cell passage and freezing stability need to be assessed. Within the same
run, QC performance (%RE ≤30%) assessed against calibration standards
in both “fresh” and ‘recently frozen’ cells.
Ranges for parameters such as assay signal, cell growth rate, and viability
should also be considered as appropriate.
New working banks established outside of validation require qualification.
Critical Reagents to be assessed: assay performance with multiple
FBS/FCS lots/sources and different media should be evaluated.
![Page 36: Outcomes of the Global Bioanalysis Consortium s ... · Outcomes of the Global Bioanalysis Consortium’s Recommendations: Large Molecule and All Molecules ... Recommended a draft](https://reader033.vdocument.in/reader033/viewer/2022060223/5f07cad67e708231d41ec4e9/html5/thumbnails/36.jpg)
Scope of review was reagents used in PK, ADA and biomarker assays.
Identified four general areas with significant guidance gaps and range of practice:
• Critical reagent documentation
• Critical reagent generated in In-house vs. obtained from a commercial source
• Changing Critical Reagents
• Stability of critical reagents
After extensive discussions, literature and global guidance review draft recommendations
which were first released in June 2012 for feedback from worldwide bioanalytical
community.
Direct comment was solicited through the use of a questionnaire and from conference
presentations, roundtables, responses from EBF, JBF and many individual companies.
The consolidated responses where used to help define the final recommendations and
best practices
Survey results determined only 50% of respondents had documentation for lot changes,
and about 50% having a procedure to address lot changes. Most surveyed indicate
having a procedure for critical reagent production but no procedure for expiry
extension.
L4: Reagents and their stability
![Page 37: Outcomes of the Global Bioanalysis Consortium s ... · Outcomes of the Global Bioanalysis Consortium’s Recommendations: Large Molecule and All Molecules ... Recommended a draft](https://reader033.vdocument.in/reader033/viewer/2022060223/5f07cad67e708231d41ec4e9/html5/thumbnails/37.jpg)
L4: Reagents and their stability Changing Critical Reagents
Changing critical reagents represents perhaps the most significant gap and challenge we identified.
Verification is typically done using QCs and criteria used for method acceptance.
The L4 Team recommends testing of new reagents in a functional assay based on a priori assay acceptance criteria. A two-tiered approach for the qualification of these critical reagents is proposed, with the level of testing being driven by the extent of change to that reagent
Examples of minor changes: new reagent lot derived from a previously qualified lot, a new affinity purification of polyclonal sera from the same animals, or a new conjugation using the same protein lot.
Examples of major changes would include antibody lots obtained from new animals, new clones for monoclonal antibody production or new cell lines for the generation of recombinant material
![Page 38: Outcomes of the Global Bioanalysis Consortium s ... · Outcomes of the Global Bioanalysis Consortium’s Recommendations: Large Molecule and All Molecules ... Recommended a draft](https://reader033.vdocument.in/reader033/viewer/2022060223/5f07cad67e708231d41ec4e9/html5/thumbnails/38.jpg)
•Recommend five levels of QC samples including the
lower and upper limits of quantitation for PK assays.
For immunogenicity assays the low positive control
close to the cut-point should be included.
•One run for single reagent with minor change.
•Recommend where possible testing in parallel with the
current or original reagent.
•Recommend monitoring instrument response from QC
samples as a tool for gauging assay performance only,
and not as an acceptance criterion.
•Results should be documented. No further validation
is required, if acceptance criteria is met.
•If acceptance criteria are not met, a single failure
might be resolved by a re-preparation of standards and
QCs and reanalysis. However, repeated failures would
indicate the need to prepare a new reagent lot, or new
reagent.
Minor Change - Qualification Major Change - Qualification
The following additional
considerations should be taken
into account for major changes
• Prior to qualification in assay,
the reagent might need to be
characterized for key
properties ( Eg. binding affinity)
•Recommend minimum of
three runs to capture the assay
performance
•If assay acceptance criteria
are not met and/or the assay
performance is altered (Eg.
changed LLOQ), but the assay
still remains fit-for-purpose, a
more extensive qualification or
partial validation may be
required
![Page 39: Outcomes of the Global Bioanalysis Consortium s ... · Outcomes of the Global Bioanalysis Consortium’s Recommendations: Large Molecule and All Molecules ... Recommended a draft](https://reader033.vdocument.in/reader033/viewer/2022060223/5f07cad67e708231d41ec4e9/html5/thumbnails/39.jpg)
The regulatory guidance states the expectation on stability shall be assured but
the guidelines do not specify how reagent stability should be assigned and
tested.
Recommendations and best practices:
Assigning test/retest rather that expiry dates to define critical reagent stability
Initial dates can be assigned based on the experimental data, information
from vendor, or experience with similar classes of reagents
Reagents stability testing runs may be included in method development,
method validation, or independent specific reagent stability testing for
intended use
Reagent expiration extension and re-qualification for regulated study support
shall be guided by a defined process which outlines testing and acceptance
criteria
Generation of large amounts of a single lot of a critical reagent reduces risk of
lot changes but requires that large amounts are stored in small aliquots often
at very low temperature to cover the anticipated life of the assay
L4: Reagents and their stability
Reagent stability assignment and testing
![Page 40: Outcomes of the Global Bioanalysis Consortium s ... · Outcomes of the Global Bioanalysis Consortium’s Recommendations: Large Molecule and All Molecules ... Recommended a draft](https://reader033.vdocument.in/reader033/viewer/2022060223/5f07cad67e708231d41ec4e9/html5/thumbnails/40.jpg)
Regulations have limited guidance for documentation of assay procedures relating to reagents
and stability. While regulations generally require documentation in the form of an SOP there are
inconsistencies and gaps in what and how to document.
Recommendations:
Documentation can take many forms; can be an SOP or similar document as determined by
your business practices
Identify critical reagents for a specific method, e.g. in the method SOP
Procedure and a priori acceptance criteria for qualification of new reagent lots and location
of this information, e.g. qualification report.
Procedure for determination of reagent stability including;
• How stability will be evaluated with a priori acceptance criteria, frequency, storage
conditions and retesting criteria (failed stability batch).
• How to document stability evaluations, e.g. separate stability batch
• Where to document stability data, e.g. stability report
Procedure to define expiry/retest date, for extending expiry/retest date with a priori
acceptance criteria.
L4: Reagents and their stability
Key Recommendations: Documentation
![Page 41: Outcomes of the Global Bioanalysis Consortium s ... · Outcomes of the Global Bioanalysis Consortium’s Recommendations: Large Molecule and All Molecules ... Recommended a draft](https://reader033.vdocument.in/reader033/viewer/2022060223/5f07cad67e708231d41ec4e9/html5/thumbnails/41.jpg)
L5: System Documentation
• Standard Operating Procedure (SOP)
• Installation Qualification (IQ)
• Operational Qualification (OQ)
• Performance Qualification (PQ) (If Significant Software Component)
User Requirements
Validation Plan
Test Worksheets
Validation Summary
Traceability Matrix
Test Incident Log
• Change Control (If Applicable)
• Configuration Only Change Control (If Applicable)
• Configuration Management
![Page 42: Outcomes of the Global Bioanalysis Consortium s ... · Outcomes of the Global Bioanalysis Consortium’s Recommendations: Large Molecule and All Molecules ... Recommended a draft](https://reader033.vdocument.in/reader033/viewer/2022060223/5f07cad67e708231d41ec4e9/html5/thumbnails/42.jpg)
• Must be done before assay validation
• Should be included in original PQ for automated system
• No need to re-validate anything already validated
• Includes integration of LIMS in process
• If module added after PQ, there are three options:
o Generate IQ/OQ/PQ documentation as defined
above for automated systems
o Generate a Change Control to another validation
o Perform as part of validation for another system
(including analysis systems)
L5: Validation of a Modular System
![Page 43: Outcomes of the Global Bioanalysis Consortium s ... · Outcomes of the Global Bioanalysis Consortium’s Recommendations: Large Molecule and All Molecules ... Recommended a draft](https://reader033.vdocument.in/reader033/viewer/2022060223/5f07cad67e708231d41ec4e9/html5/thumbnails/43.jpg)
L5: Accuracy and Precision Testing
• Scenario 1: Manually validated assay (benchmark
only, out of scope of committee)
• Scenario 2: Assay validated with robotics
• Scenario 3: Manually validated assay along with
assay validation with robotics
• Scenario 4: Manually validated assay followed by
assay validation with robotics LATER
![Page 44: Outcomes of the Global Bioanalysis Consortium s ... · Outcomes of the Global Bioanalysis Consortium’s Recommendations: Large Molecule and All Molecules ... Recommended a draft](https://reader033.vdocument.in/reader033/viewer/2022060223/5f07cad67e708231d41ec4e9/html5/thumbnails/44.jpg)
L5 : Automation Practices in Large
Molecule Bioanalysis
System Documentation
SOPs
IQ/OQ/PQ
Change Control
Configuration Management
Validation of a Modular System
Accuracy and Precision Testing
Multiple scenarios exist depending on the assays
![Page 45: Outcomes of the Global Bioanalysis Consortium s ... · Outcomes of the Global Bioanalysis Consortium’s Recommendations: Large Molecule and All Molecules ... Recommended a draft](https://reader033.vdocument.in/reader033/viewer/2022060223/5f07cad67e708231d41ec4e9/html5/thumbnails/45.jpg)
L6 - Immunogenicity (effect on PK)
In an upcoming white paper we will provide consensus based
recommendations that are supported by case studies:
• Prior to the development of PK assays the bioanalytical scientist should
consider how the methods may possibly be made ADA tolerant.
• We would recommend the testing of the PK assay for antibody tolerance, but
the evaluation should be done based on potential risk.
• A risk-based approach should be used in conducting investigations, i.e. not
all abnormal PK results require a thorough investigation.
• It is advisable to consider the expected drug therapeutic window during drug
PK method development to gauge the appropriate amount of effort when
examining ADA impact on assay performance.
• PK and ADA data should be considered in combination with other PD and
efficacy markers.
• We advocate covariate PK data analysis that takes into account the effects of
clinically-relevant parameters.