Topic 1:

Driving innovation in Discovery Bioanalysis through Robotics and Automation of Data Workflows due to the emergence of several complex/mixed modalities and novel targets: Modernization by modular automation to improve throughput, robustness, reproducibility and reduce hands-on time requirements; Need to incorporate emerging analytical instrumentation, High Throughput Mass Spectrometry, Next-Generation Sequencing (NGS); Automation of data analysis, review by exception and facilitation of Emerging AI/ML Approaches.

Topic 2:

Opportunities & Challenges in using Artificial Intelligence/Machine Learning (AI/ML) for ADME and Discovery Bioanalysis and potential future expansion in Regulated Bioanalysis by enhancing drug discovery design and efficiency, data readiness to ensure quality; Using AI/ML to design better compounds, Predict ADME endpoints, optimize absorption and clearance, to predict Non-Regulated BA methods, fine tune CYP3A4 contribution for early human dose projection; Accelerate timelines and produce high-quality data. 

Topic 3:

Human Radiolabeled Mass Balance Studies and the Implications to Regulated Bioanalysis: What are they and where do the fall in drug development progression? The 2024 US FDA Guidance for Human Radiolabeled Mass Balance Studies; Changes and/or clarifications by guidance regarding human ADME design and conduct; When to consider nontraditional paradigms; Considerations for newer and alternative technologies and clinical designs and their application to hADME studies. 

Topic 4:

Proposals to Better Rationalize the use Chiral Assays and Accelerator Mass Spectrometry (AMS) in  support of Clinical Programs: Is it still necessary to continue monitoring the enantiomers if the pair ratio was determined to be constant after in vivo measurements? Can ADME, Mass Balance and Absolute Bioavailability be obtained in one micro tracer study? Is AMS fit-for-purpose Method Validation based on ICH M10 Guidelines? What is the best practice for FFP validation & implementation of LC-AMS?

Topic 5:

Innovative Approaches for Developing Bioanalytical Methods to measure Isobaric Saccharides as Biomarkers: Quantification of rhamnose, erythritol, lactulose and sucralose in human urine and plasma to support an indomethacin challenge study for intestinal permeability assessment in certain immunoinflammatory diseases; Technical considerations MS Tune Settings and Mobile Phase Selection; General concepts for broadly applying RPLC-MS/MS methods to other saccharides to provide confidence in disaccharide to monosaccharide ratio.

Topic 6:

Comparing to Conventional Denaturing LCMS Techniques with native LCMS methods as alternative to address emerging challenges in Biotransformation of Alternative Antibody Formats and Transgene Product; 2D native LC-MS approach enabling direct and immunoprecipitation-free analysis of serum/plasma samples, demonstrating great potential in transgene product characterization in Gene Therapy and biotransformation study of mAbs reducing method development time and analytical bias.

Topic 7:

Continued Expansion of the use Hybrid Assays (IA-MS) in Immunogenicity: Development of a hybrid assays to assess the presence and semi-quantification of ADAs as complementary test for the understanding of results from the conventional approaches; Mass Spec ability to isotype the ADAs IgM and IgG (with IgGs subclasses) in the same extraction of clinical samples; Development of a dual immunocapture to detect and semi quantify IgE ADAs specific to the drug.

Topic 8:

Challenges in Measuring Post-Translational Modifications (PTMs) by Hybrid Assay (IA-MS): Development Strategy to Quantify Site-Specific Phosphorylated Protein Biomarkers (phosphorylation, glycosylation, acetylation, methylation, ubiquitination, nitrosylation) for Clinical Development; Overcoming Mass Spec insufficient sensitivity, reproducibility, and throughput for PTMs quantification in clinical applications; Novel anti-phosphopeptide immunocapture strategies for sensitive quantitation of multiple site-specific phosphorylated protein biomarkers for clinical development.

Topic 9:

Lesson Learned form the development of Hybrid Assays (IA-MS) for Glycopeptide PK Monitoring for a Therapeutic Fusion Protein with Multiple Glycosylation Sites: Complex composition of therapeutic glycoproteins and glycosylation characterization; Extensive HRMS identification of glycopeptides; Use of micro-LC-MS/MS in a trap & elute format; Novel approaches to assess individual glycoform PK profile and its contribution to the overall PK properties of glycoprotein.

Topic 10:

Recent Developments in Intact IA-MS Analysis of Symmetric High Mannose (on both heavy chains) vs Asymmetric High Mannose (only on one heavy chain) vs other Glycoforms of monoclonal Antibody (mAb) to assess systemic clearance; Novel enzyme digestion to shorten glycans which can be implemented in routine bioanalysis; Needs to determine symmetric/asymmetric high mannose content in addition to total high mannose content as part of NBE drug product characterization.

Topic 11:

Unique Technical Advantages and Current Challenges to Quantify Endogenous Protein Biomarkers in Tissue by Mass Spec:  High specificity for proteoforms and bioanalytical accuracy maintained in complex matrices vs Issues related to sample preparation, assay calibration, and normalization in Tissue; Overcoming sensitivity challenges due to low expression level or trace abundance post-translationally modified form of the target protein by modern Mass Spec platforms with sensitivity in pg/mL levels.

Topic 12:

Advanced applications of Hybrid Assays (IA-MS) Protein Bioanalysis for Bioavailability and Site-of-Action Target Engagement: Measurement of the VHS variant to biotherapeutic ratio to demonstrate that VHS variant bioavailability was comparable to the biotherapeutic; Comparing Target Engagement (TE) data from Fresh Frozen Tissues with FFPE Tissues; Importance to obtain high sensitivity protein data from FFPE tissues since the vast majority of patient tissues are stored as FFPE.

Topic 13:

Increasing need to Measure Tissue Specific PD Biomarkers in oncology and immunology clinical studies: Pros/Cons of  Immunohistochemistry (IHC) vs RNA-based vs Mass Spec-based methods to measure biomarkers in biopsy; Considerations on how sample collection may affect what types of assays might be employed; Benefits and limitations of tissue biopsies collected at the clinical site fixed in paraffin (FFPE) or Flash Frozen and analyzed in single-plex or multiplexed. 

Topic 14:

Tips, Tricks, and Strategies for Improving the Quantification of Protein Biomarkers by Mass Spec in Tissue: Comparing and contrasting Tissue Quantification Normalization by tissue weight, protein content, normalization peptides and effect on data; Considerations on adding SILAC labelled protein to tissue for normalization; Effect of blood contamination on PK and PD measurements; Comparing different lysis buffers and manual/automated sample preparation workflows and their efficiency.

2025 White Paper on Mass Spectrometry, Chromatography, and Sample Preparation

Consensus & Conclusions on Mass Spectrometry, Chromatography, and Sample Preparation for 2025 White Paper