2019 White Paper on Recent Issues in Bioanalysis: FDA Immunogenicity Guidance, Gene Therapy, Critical Reagents, Biomarkers and Flow Cytometry Validation (Part 3 – Recommendations on 2019 FDA Immunogenicity Guidance, Gene Therapy Bioanalytical Challenges, Strategies for Critical Reagent Management, Biomarker Assay Validation, Flow Cytometry Validation & CLSI H62)

Abstract

The 2019 13th Workshop on Recent Issues in Bioanalysis (WRIB) took place in New Orleans, LA, USA on April 1–5, 2019 with an attendance of over 1000 representatives from pharmaceutical/biopharmaceutical companies, biotechnology companies, contract research organizations and regulatory agencies worldwide. WRIB was once again a 5-day, week-long event – a full immersion week of bioanalysis, biomarkers, immunogenicity and gene therapy. As usual, it was specifically designed to facilitate sharing, reviewing, discussing and agreeing on approaches to address the most current issues of interest including both small- and large-molecule bioanalysis involving LCMS, hybrid LBA/LCMS, LBA cell-based/flow cytometry assays and qPCR approaches. This 2019 White Paper encompasses recommendations emerging from the extensive discussions held during the workshop and is aimed to provide the bioanalytical community with key information and practical solutions on topics and issues addressed, in an effort to enable advances in scientific excellence, improved quality and better regulatory compliance. Due to its length, the 2019 edition of this comprehensive White Paper has been divided into three parts for editorial reasons. This publication (Part 3) covers New Insights in Biomarker Assay Validation, Current & Effective Strategies for Critical Reagent Management, Flow Cytometry Validation in Drug Discovery & Development & CLSI H62, Interpretation of the 2019 FDA Immunogenicity Guidance and Gene Therapy Bioanalytical Challenges. Part 1 (Innovation in Small Molecules and Oligonucleotides & Mass Spectrometry Method Development Strategies for Large Molecule Bioanalysis) and Part 2 (Recommendations on the 2018 FDA BMV Guidance, 2019 ICH M10 BMV Draft Guideline and regulatory agencies' input on bioanalysis, biomarkers, immunogenicity and gene therapy) are published in volume 11 of Bioanalysis, issues 22 and 23 (2019), respectively.

Insights on Droplet Digital PCR–Based Cellular Kinetics and Biodistribution Assay Support for CAR-T Cell Therapy

“…In the drug discovery and development phase of in vivo and ex vivo gene therapy, cellular kinetics and biodistribution have become an integral part of efficacy and safety assessment in nonclinical and clinical studies (24). The emergence of the ddPCR technology over the past ten years has expanded the opportunity and potential for its application toward the innovative in vivo and ex vivo gene therapy regardless of therapeutic areas (i.e., immuno-oncology cell therapy and gene therapy in rare diseases).…”

Flow cytometric method transfer: Recommendations for best practice

“…The SOP should include a complete description of the method including the instrument setup procedures, reagents (vendors and catalog number, clone and fluorophore conjugation for monoclonal antibodies), sample processing, data review, run acceptance criteria and reporting. The transfer plan should include a description of the experiments to be performed including sources of the samples, number of replicates, statistical evaluation and acceptance criteria (Cossarizza et al, 2019; der Strate et al, 2017; Piccoli et al, 2019; Selliah et al, 2019). Aforementioned recommendations are also consistent with CLSI Guideline H62: Validation of Assays Performed by Flow Cytometry (CLSI, 2021).…”

2020 White Paper on Recent Issues in Bioanalysis: BMV of Hybrid Assays, Acoustic MS, HRMS, Data Integrity, Endogenous Compounds, Microsampling and Microbiome ( Part 1 –Recommendations on Industry/Regulators Consensus on BMV of Biotherapeutics by LCMS, Advanced Application in Hybrid Assays, Regulatory Challenges in Mass Spec, Innovation in Small Molecules, Peptides and Oligos)

“…The most prominent vector is AAV, which is particularly suited to infect both dividing and nonor slowly-dividing cells. Initial recommendations on the use of hybrid assays for gene therapy have already been issued in the 2018 White Paper in Bioanalysis and sustained in 2019 White Paper in Bioanalysis [20,25]. It was agreed that hybrid assays are becoming a critical tool not only for quantifying but also characterizing the transgene products for which specificity is key that would not be easily available from other types of assays.…”