on behalf of the EBF EBF Training day...Update on the guidelines and current practice in industry...

Update on the guidelines and current practice in industry

Michaela Golobon behalf of the EBF

EBF Training dayCritical Reagents for LBA

Altis Grand Hotel, Lisbon - 14 May 2018

http://www.europeanbioanalysisforum.eu

Reagents

ØClassical LBA reagents are– mAbs– pAbs– Recombinant proteins (receptors, ligands,…)– Peptides – Oligonucleotides, Aptamers– Conjugated reagents– …

http://www.europeanbioanalysisforum.eu 2

„Critical reagents“ - What does this mean?


If a change in the reagent has an impact on your assay performance & outcome it is a critical reagent.

Critical Reagents


CR can be different depending on assay format & what the assay is used for!

LBA indirectly measures the binding interactions between the analyte of interest and the assay reagents.

Assay quality relies on reagent quality!

Critical Reagent


Before you start

Þdefine your assays critical reagents!

1. Scientific question: what reagent is critical for what measurement (free/ total, interference in matrix, species dependent/independent)

2. How can I ensure equal quality and long-term performance of LBA => life-cycle management

What is regulated and how?


ØEMA Guideline on bioanalytical method validation (2012)EMEA/CHMP/EWP/192217/2009 Rev. 1 Corr. 2** )

ØFDAGuidance for Industry Bioanalytical Method Validation (2001 & 2013 Rev1 (draft))

ØMHLW Guideline on Bioanalytical Method (Ligand Binding Assay) Validation in Pharmaceutical Development (2014)

EMA (2012)


Ø 7.1.1.12. Reagents

Critical reagents, including binding reagents (e.g. bindingproteins, aptamers, antibodies or conjugated antibodies) andthose containing enzymatic moieties have direct impact on theresults of the assay and therefore their quality must be assured.Accordingly, when changing reagent batches during validation orsample analysis the analytical performance of the method mustbe verified to ensure that it is not altered compared with theoriginal or previous batch. Conditions guaranteeing themaintenance of the stability of both non critical reagents (e.g.buffers, diluents or acidification reagents) and more importantlyof the critical reagents should be documented in order to ensurethat the performance of the method is not affected over time.

FDA (2001)


Key reagents, such as antibody, tracer, reference standard, and matrix should be characterized appropriately and stored under defined conditions.Assay reoptimization or validation may be important when there are changes in key reagents, as follows:

Ø Labeled analyte (tracer) - Binding should be reoptimized.- Performance should be verified with standard curve and QCs.

Ø Antibody- Key cross-reactivities should be checked. - Tracer experiments above should be repeated.

Ø Matrix- Tracer experiments above should be repeated.

FDA (2013, draft)


Ø Key Reagents Key reagents, such as reference standards, antibodies, tracers, and matrices should be characterized appropriately and stored under defined conditions. Assay reoptimization or validation may be important when there are changes in key reagents. For example:

Ø Labeled analytes (tracers) - Binding should be reoptimized. - Performance should be verified with standard curve and

QCs. Ø Antibodies

- Key cross-reactivities should be checked.- Tracer experiments above should be repeated.

Ø Matrices - Tracer experiments above should be repeated.

MHLW (LBA 2013)


Ø 6.5. Critical reagent Critical reagents are usually binding reagents (labeled or unlabeled antibodies) that have a direct impact on the results of ligand-binding-based bioanalyticalmethods. A critical reagent should be selected by considering the specificity for the analyte and stored under conditions that ensure consistent quality. The quality of critical reagent should be appropriately maintained throughout the period of use in analytical method validation and study sample analyses. Partial validation is required when the critical reagent lot is changed.

MHLW (LBA 2013)


Definition: Critical reagentA reagent that has a direct impact on the results of an LBA-basedbioanalytical method, an example being binding reagents (e.g., unlabeled or labeled antibodies).

Q&A:

Critical reagents >>(Q25). Should an expiration date be established for critical reagents?(A25). Expiration date is not necessarily required for a critical reagent, as long as the quality is ensured by evaluation of data from calibration standards and QC samples during the period the reagent is used.

ICH M10:Weehawken/Lisbon Sister Workshop, Sept. 2018


ICH M 10 – recommendations from WeehawkenWorkshop

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ØCritical ReagentsIdentify critical reagents, normally the capture and detection antibodies, in the bioanalytical method and manage them appropriately to ensure consistent analytical performance of the method.


ICH M 10 – recommendations from LisbonWorkshop

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Critical Reagents RecommendationsØ Critical reagents should be identified and defined in the assay

methodØ Should be monitored through the life cycleØ Assess for lot to lot changes and consider the impact of the

change on the assayØ Documentation: source and origin by CoA or technical

datasheet


Best practice (2014) – Global Bioanalysis Consortium (GBC) L4


GBC L4 => Focus


Management of critical reagents for pharmacokinetic, immunogenicity and biomarker ligand binding assays

Reagents are the foundation of LBA assays; specificity, selectivity and sensitivity of LBAs are dependent on reagents

GBC L4 => main question


What is a Critical Reagent for my assay?ØNot only based on specific or rare reagents

like: „reagents difficult to make, replace, acquire or substitute

ØAlso more generic reagents i.e. AB againstlight or heavy chain antibodies can be criticalin some assays

The need to define a reagent as critical demands active management of reagent

availability and reproducibility.

GBC L4 => recommendation


Bioanalytical groups should have an a priori plan or framework for LBA critical reagent management. This plan should Ø Identify critical reagents and risks to resupply and lot

changes, whether provisioned internally or from a commercial source

Ø Have a scientifically justified plan to address lot changes and stability of critical reagents, and

Ø Define critical reagent-related documentation.

GBC L4 => CR life cycle management


O‘Hara & Theobald (2013) : Life Cycle management of CR


A review article highlighting requirements for life cycle management of LBA critical reagents

Ø reagent generationØ characterizationØ resupplyØ inventoryØ stability

O’Hara DM, Theobald V. Life cycle management of critical ligand-binding reagents. Bioanalysis 5(21), 2679–2696 (2013).

Wang et al. (2015)requirements to assess analyte and reagent

stability

Wang J, Nowatzke W, Ma M.Current industrial practices and regulatory requirements to assess analyte and reagent stability using ligand-binding assays.Bioanalysis. 2015;7(11):1371-84. Review


A review article highlighting current best practices to evaluate analyte and reagent stability using LBA in the regulatory setting.

Paper compares :• expectations from US, EU and Japan on stability

assessments• Industrial practices of stability including reagent

stability and documentation requirement

Acknowledgment

ØCurrent EBF Reagents Team– Susanne Pihl– Barry van der Strate– Birgit Jaitner– Laurent Vermet

ØAll former members of the critical reagent team

ØEBF community


on behalf of the EBF EBF Training day...Update on the guidelines and current practice in industry...

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