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By Faye Vazvaei, Linzhi Chen, and Hao Jiang

Faye Vazvaei-finalLin Chen-finalHao Jiang-finalImmunogenicity testing is required for biologic drug development, as well as for biosimilars approval, in order to understand the impact of the patient immune response on drug safety, pharmacokinetics, and efficacy. Traditionally, ligand-binding assays are used for immunogenicity testing either by inferring the presence of antidrug antibodies (ADAs) using a bridging assay format (drug as the capture and detection reagents) or a direct assay format (capturing with the drug and detecting with an anti-species antibody). Usually, a tiered approach (screen, confirm, and titer) follows, providing a positive or negative result for the presence of ADAs and an associated titer value for positive samples. In the past few years, liquid chromatography/tandem mass spectrometry (LC/MS/MS) has risen into a widely accepted technology platform for quantitative bioanalysis of biotherapeutics and biomarkers, mainly due to its superior specificity, selectivity, and a wide dynamic range. When combined with immune-affinity sample cleanup and enrichment, it can also deliver sensitivity comparable to traditional ligand binding assays. As the use of this hybrid approach for protein quantitation by LC/MS/MS is maturing, it has also been explored for other applications such as investigating for the presence of ADAs and for determination of their isotypes and subclasses within a single assay.

The sunrise session LC/MS/MS ADA Analysis at the 2015 AAPS National Biotechnology Conference is intended to showcase the recent progress in ADA analysis using LC/MS/MS. Linzhi Chen, Ph.D., from Boehringer Ingelheim Pharmaceticals and Hao Jiang, Ph.D., from Bristol-Myers Squibb Co. are among the leaders in this field, and they will present their research in this new frontier.

The first speakers will present a novel approach using LC/MS/MS for simultaneous ADA isotyping and semi-quantitation. The key to this approach is identifying and using unique yet universal peptides for each isotope/subclass, including all allotypes. Preliminary results show that this approach is feasible for immunogenicity assessment. A similar approach will be presented by the second speaker with a focus on simultaneous measurement of residual monoclonal antibody drug, residual endogenous IgGs, and ADA positive control to assist the optimization of a neutralizing antibody (Nab) assay utilizing bead extraction and acid dissociation. By applying this approach, a few critical questions during cell-based assay development have been successfully answered for several immunogenicity projects. Therefore, this useful LC/MS/MS approach has been found invaluable as a complementary technique in immunogenicity evaluation.

This session will show how LC/MS/MS may be utilized to address questions on immunogenicity assessment and overcome challenges such as ADA isotyping. As this technique becomes more mature in ADA analysis, it will find its place as an orthogonal approach for ADA assessment. The session is recommended for scientists engaging in ADA and immunogenicity determination and evaluation.

We look forward to seeing you there!

Faye Vazvaei is a senior principal scientist at the DMPK and Bioanalytical R&D Department of Roche Innovation Center New York. She currently serves as the AAPS Bioanalytical Focus Group (BFG) Chair.
Linzhi Chen, Ph.D., has been with Boehringer Ingelheim Pharmaceuticals for the past 14 years. His recent research focuses on developing immunopurification-LC/MS for intact protein analysis, protein quantitation via digestion and anti-drug antibody analysis.
Hao Jiang is a senior research investigator II at Analytical and Bioanalytical Development of Bristol-Myers Squibb Co. His current research focus is protein bioanalysis for pharmacokinetics, immunogenicity, and biomarkers by LC-MS.