Topic 1:

Novel, Streamlined, and Fully Automated Approaches for High Throughput Clinical LBA PK and ADA Development: Reagent Conjugation using the Big Tuna instrument; Screening of monoclonal Antibodies for PK assay development; Concurrent ADA methods development for pre-existing antibody screening in support of final drug candidate identification using clinical ADA assay platform; Tips and Tricks when considering using Automation; Benefits of consistent method development approaches/testing/data assessment across multiple projects.

Topic 2:

Advanced Application of DOE to improve LBA Robustness and Ruggedness: Use of 15 Test Factors based on upfront review and ranking of development data, scientific experience, and commonly expected sources of variability; Testing the factors in  16-run Resolution III design and reducing the total number of runs needed compared to a traditional checkerboard design;  Impact of plate manufacturer with interaction of coating concentration and time as critical robustness factors. 

Topic 3:

Challenges and Lessons Learned on Selecting Appropriate Matrix and Matrix Handling Conditions for monoclonal Antibodies and other Protein Targets for PK and PD LBA; Effect of blood processing on the target of interest, whether platelets contain the target of interest and the freeze/thaw stability of endogenous targets and recombinant proteins; Platelet degranulation resulting in an artifactual elevation in a mAb drug target.

Topic 4:

Lesson Learned for LBA Cross-validation for PK assays and for immunogenicity of biotherapeutics ADA/NAb assays; Updated Considerations for Cross-Validating Different Bioanalytical Methods, including method modifications to fully validated assays, and cross-validating multiple bioanalytical laboratories; Approaches taken to address failed cross-validations after study samples have already been tested; Sample size considerations, use of QC vs. incurred study samples, criteria for result comparison.

Topic 5:

Novel Strategies for Complex Drug Modalities ADA Assay: What are the Regulatory Expectations regarding ADA method assay format, number of replicates and validation? Considerations on new and challenging drug modalities. How have methods evolved to meet these challenges? Implications for ADA method development, validation and sample testing for a complex modality and how the method is used to understand safety risks.

Topic 6:

Due Diligence for Cut-point Establishment of a Plate-based NAb Assay: CBA not feasible for the Nab method based on therapeutic target and MoA; Existing screening, confirmatory, and titering assays have sample treatment steps that were foundational for the Nab method development; Importance of diligence activities, including competitive method phase optimization and proper statistics, underscore the validity of an unusually low method cut point.

Topic 7:

Justification of Utilizing Bridging LBA Format instead of Cell-based Assay (CBA) to assess anti-CAR total binding antibodies; Unique considerations of assay design and method optimization for the assessment of ADA to bispecific CAR-T therapy;      Addressing challenges related to the high incidence of pre-existing ADA against CAR protein during method development; Strategy to determine cut point and key assay parameters in the presence of pre-existing ADA during method validation.

Topic 8:

How to Remove IgM Interference in LBA? - A novel approach: Mitigating Rheumatoid Factor Interference by IgM-specific proteases by preventing false positives and improving assay accuracy; Implementing IgM digestion in ADA Assays simplifies IgM isotyping and enhances the detection of IgG ADAs; Protease treatment can be easily integrated into existing assays without compromising throughput or reliability, applicable to both clinical and preclinical settings, particularly in biologics development. 

Topic 9:

Consideration on NUcleic acid Linked Immuno-Sandwich Assay (NULISA) as new Emerging Technology for ultrasensitive detection of analytes in biological matrices: Comparison of cross platform results from NULiSA as compared to Olink for Multiplex and MSD and Simoa for single-plex; Where was good correlation seen and where did results differ? Does NULiSA fill some of the gaps seen with the OLink in terms of sensitivity?

Topic 10:

Unbiased Evaluation of NULISA beyond Biomarkers & Proteomics applications: Unconventional NULISA assessment for PK Determinations due to its remarkable sensitivity, broad dynamic range and full automation capabilities; A PK assay with a broad dynamic range developed on the NULISA platform generated a standard curve of >6 dynamic range using the same capture and detection antibody pairs used in the Gyrolab version of the assay. 

Topic 11:

Multiplex Vaccine LBA: Striking a balance to improve Throughput while achieving Sensitivity across Analytes: Needs for multiplex LBA  due to the limited availability of matrix for pediatric clinical trials; Considerations on the two main multiplex technologies  Luminex and MSD, elucidate the benefits and challenges of multiplexing; Multiplexing 15-32 serotypes, a challenging feat to balance the performance of that many analytes when multiplexed in a single well.

Topic 12:

Unique approaches used to Generate Novel Critical Reagents when the "tried and true" approaches failed: Established protocols for reagent conjugations vs new modalities complexity  and assays used to measure them; Using novel Ruthenium "motorcycle sidecar" conjugation approach for enhanced MSD LBA sensitivity; Using  site-specific reagent conjugation approaches when random labeling procedures fail; Using  VLP's (virus-like particles) as capture reagents for difficult cell membrane target proteins to maintain native folding structure.

Topic 13:

Using Modeling to Predict Stable Formulations for scFv-Fc Conjugated Critical Reagents to Support CAR-T Clinical Programs: Biotinylated and ruthenylated scFv-Fc critical reagents generated using platform conjugation and formulation conditions precipitated after freeze thaw; In silico modeling and confirmatory experimental range finding studies enabled the development of stable formulations for the critical reagents. The protein concentration, pH, and excipients of the formulation buffer all contribute to the stability of the critical reagents.

Topic 14:

State-of-the-art Approaches for Screening Critical Reagents to support both ADA and NAb Assay: Affinity-Maturation Engineering via phage display to optimize anti-idiotypes antibodies; Recombinant Anti-IDs Generation via Hybridoma Platform and Kinetic measurements using Octet; Using functional assay to Confirm Improved Anti-IDs are fit-for-purpose critical reagents; How to develop critical reagents for domaining assay for tri-specific mAbs, especially for scFV arm.

Topic 15:

Importance of Performing a Thorough Characterization of Critical Reagents to develop Robust Bioanalytical LBA: Stress Studies as crucial step for a more Comprehensive Understanding of Ligand Stability:  Need for further purification to ensure high purity of the reagent and performance consistency; Extended characterization of ligands needed to evaluate oligomerization and post translational modifications (PTM) affecting the ligand binding to the drug.

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

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