Topic 1:

Challenges, Pitfalls, Novel Approaches in the Mass Spec Bioanalysis of Lipid Conjugates siRNAs: Improved siRNAs biodistribution & stability by covalent conjugation with cholesterol or fatty acids; Significant impact of conjugation on extraction and chromatography,  Re-engineering of SPE and ion pairing chromatography for simultaneous detection of both strands and to avoid chromatographic artifacts; Use of LCMS with PNA probes & µflow chromatography to increase the assay sensitivity.

Topic 2:

Efficient Strategies for Quantification of double-stranded siRNA Deaminated Metabolites to overcome the complications introduced by deamination; Quantification siRNAs with a single deamination site by Mass Spec method provides specificity to differentiate parent and deaminated metabolites; However, quantification of siRNA with multiple deamination sites is more complicated, and specific LCMS strategies need to be considered to quantify such siRNA; FFP Validation to meet accuracy/precision criteria in monkey studies.

Topic 3:

Mass Spec as the Most Selective Platform for Quantifying siRNA and its truncated metabolites: Considerations on Hybridization ELISA (HELISA) high sensitivity and throughput but lacks selectivity and cannot differentiate between the parent siRNA and its truncated metabolite(s); Comparison of HELISA and LCMS employed to measure siRNAs concentrations in TK and PK studies; Crucial importance to bridge the two platforms to fully utilize the available data for understanding PK-PD.

Topic 4:

Innovative Bioanalytical Strategies for Antibody-Oligonucleotide Conjugates (AOC) by Mass Spec and LBA: Why the need to develop antibody-oligonucleotide conjugates? Understanding AOC different design, advantages and bioanalytical challenges for different components AOC, including antibody-oligonucleotide conjugate, total and free antibody, and free oligonucleotide; Using LBA for quantification for total antibody and free antibody and hybridization LC-MS/MS for unconjugated (free) oligonucleotide.

Topic 5:

Streamlined Decision-making process on which ADC Related Bioanalytical Assays need to be Validated based on early discovery data as well as current Regulatory Guidance; Mass Spec or LBA? Deciding which ADC assays should be validated by Mass Spec or LBA based on critical reagent availability and timelines; Performing early feasibility test to ensure Mass Spec-based assays will be successfully validated.

Topic 6:

State-of-the-art Bioanalytical Strategies for Antibody-drug conjugates (ADC) by Hybrid Assays (IA-MS) used in various development stages, especially in discovery setting for ADC lead selection: Technical considerations for method development and fit-for-purpose BMV; Screening candidate lead ADC molecules during discovery mouse PK to support portfolio program development; Investigating new linker & payload chemistries to support platform program for future ADC development.

Topic 7:

How did the development of ADC assays in Regulated Bioanalysis shape Bicycle Toxin Conjugates (BTC) quantitation? What do ADC & BTC methods have in common? Unique BTC strategies employed for success in routine quantitation to meet Regulatory Guidance and robust clinical performance;  Necessary selective sample preparation and extraction techniques for accurate and reproducible payload determinations; Pros/Cons of QQQ vs HRMS for BTC and challenges experienced during method development.

Topic 8:

Enabling Bioanalysis for Complex Large Molecule with Mass Spec Immuno-Enrichment (IE) and/or Immuno-Depletion (ID): Use of cost-effective IE Mass Spec to quantify a mouse monoclonal antibody in mouse plasma; Combining both IE and ID sample preparation techniques in the bioanalytical workflow to investigate the observed payload discrepancy of an ADC during its plasma stability assessment; T-cell engager bioanalysis by leveraging the unique selectivity ID/IE Mass Spec.

Topic 9:

Present challenges and recent progresses in Mass Spec Multiplexing Assay Development and Validation: Using Multiplex LCMS assays for exploratory measurements of Neurological indications including Parkinson's disease and Alzheimer's and validation considerations; What is the current state on Regulatory Acceptance of LDTs, RUO assays that utilize HRMS as a detector (Orbitrap & QTOF)? Are there special considerations for using Seed amplification assays (SAAs) in clinical trials? 

Topic 10:

New Progresses in Multiplexed Assay Design for Reagent-free Multi-dimensional, SEC, nano-LC HRMS to measure low-abundance peptide hormones: Advantages of using of Size-exclusion Chromatography (SEC) to replace Immuno-Enrichment (IE) for simultaneous quantitation of dissimilar peptides; Multiplexed assay for intact measurement of dissimilar peptide hormones without antibody capture; Potential for peptide hormone panel assays like the steroid hormone panels now finding way into Clinical Diagnostic labs.

Topic 11:

Assay Performance Comparison between LBA and LCMS Assays for Neurodegenerative Diseases Biomarkers:  Novel Biomarkers for Alzheimer's Disease for diagnosing, identifying the disease stage and PD endpoints; Mass Spec and Ligand-binding Assays available for plasma pTau217 biomarker and their validation; Concordance and discordance between Mass Spec and LBA data and interpretation; How the data can assist in Alzheimer's Disease diagnosis and clinical trial design.

Topic 12:

Is CSF & Urine Non-specific Binding still a Concern for Bioanalysis? Challenges of using additives in sample collection or transfer tubes to prevent non-specific binding; Number of experiments needed to be performed to ensure samples remain uncontaminated, stable during collection and storage, reliability of the sample integrity, and meet required standards for various parameters. Considerations on additives not compatible for multiple analytes in the same assay.

Topic 13:

New considerations emphasizing the Importance of Evaluating the Chemical Structure of the Analyte during Method Development to avoid issues during Sample Analysis: Struggles with the bioanalysis of an ADC-payload containing a lactone; Comprehensive evaluation of lactone hydrolysis and cyclization of carboxylate in aqueous solution and in serum at different pH to design the extraction protocols for LLE and SPE to ensure recovery of all forms to the payload in clinical samples.

2025 White Paper on Mass Spectrometry Assays - Latest Advances, Challenges and Solutions from Internationally Recognized Key Opinion Leaders

Consensus & Conclusions on Mass Spectrometry Assays - Latest Advances, Challenges and Solutions from Internationally Recognized Key Opinion Leaders for 2025 White Paper