2018 White Paper on Recent Issues in Bioanalysis: Focus on Immunogenicity Assays by Hybrid LBA/LCMS and Regulatory Feedback (Part 2 – PK, PD & ADA Assays by Hybrid LBA/LCMS & Regulatory Agencies’ Inputs on Bioanalysis, Biomarkers and Immunogenicity)

Abstract

The 2018 12th Workshop on Recent Issues in Bioanalysis took place in Philadelphia, PA, USA on April 9–13, 2018 with an attendance of over 900 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 and immunogenicity. 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 and LBA/cell-based assays approaches. This 2018 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 2018 edition of this comprehensive White Paper has been divided into three parts for editorial reasons. This publication (Part 2) covers the recommendations for PK, PD and ADA assays by hybrid LBA/LCMS and regulatory agencies’ input. Part 1 (LCMS for small molecules, peptides, oligonucleotides and small molecule biomarkers) and Part 3 (LBA/cell-based assays: immunogenicity, biomarkers and PK assays) are published in volume 10 of Bioanalysis, issues 22 and 24 (2018), respectively.

IQ Consortium Perspective: Complementary LBA and LC–MS in Protein Therapeutics Bioanalysis and Biotransformation Assessment

“…Integrated bioanalytical approaches involving LBA, MS and hybrid assays have become increasingly common as large molecule drugs have become more complex including fusion proteins, bispecifics, conjugated proteins and proteins containing non-native amino acids (Table 2) [1,2]. In case of fusion proteins, it is critical to understand the in vivo stability of the protein subunits, as unexpected proteolysis could result in a loss of potency and ultimately reduced efficacy [25].…”

“…In recent years, MS-based approaches like hybrid immunoaffinity (IA) LC-MS have been established as an alternative to traditional ligand-binding assays (LBA) for the bioanalysis of biotherapeutics [1,2]. This use has been driven largely by increasingly complex large molecular modalities in the biopharmaceutical industry pipelines, for example, therapeutic proteins, monoclonal antibodies (mAbs), bispecific antibodies, antibody drug conjugates and fusion proteins.…”

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)

“…Hybrid Assays for AAV Gene Therapy & Extracellular Vesicles: Advanced Applications Since this topic was discussed in the 2018 White Paper in Bioanalysis [20], what is the current applicability of hybrid assays in clinical and preclinical settings for gene therapy drug development? What experience exists across industry in the analysis of transgene protein expression by LCMS aiding gene therapy development?…”

“…High Resolution Mass Spectrometry for Protein Therapeutic Bioanalysis: Current Developments Have there been any advancements for sensitivity improvements following the 2018 and 2019 White Paper recommendations [20,23]? Have there been any advancements on other fronts, such as sample preparation, chromatography, native intact MS bioanalysis, or software?…”

“…• Duplicate analysis, typically used as part of LBA methods, was not considered required if an internal standard is used [5] • To ensure hybrid assay performance and robustness during the sample analysis, it was recommended that automation be used to increase reproducibility and throughput • Additional assay performance testing may also be needed when the assay is applied to new patient populations or matrices • When biotherapeutics are analyzed by hybrid assays, bridging studies should be performed between discovery and development to ensure interpretation of the results remains consistent as the method evolves [20] Hybrid Assays for AAV Gene Therapy & Extracellular Vesicles: Advanced Applications Gene Therapy…”