Topic 1:

What is the best Whole blood (WB) Preservation method for High-Parameter Flow Cytometry? Pros/Cons of commercially available stabilizers and efficacy in maintaining immune cell populations representative of fresh whole blood: Cyto-Chex, TransFix, Smart Tubes, Cytodelics; Considerations on whether WB preservation is influenced by antibody clones, fluorochromes, other components within the panel, interactions with stabilizers; Establishing effective protocols designed to minimize sample handling, maintain cellular integrity, and mitigate cell loss.

Topic 2:

Recent developments in Flow Cytometry Extended Sample Stability preserving blood or other sample matrixes for longer periods before analysis, while maintaining the integrity and reliability of the measured Cellular Markers; Cryopreservation as alternative using PBMC, frozen blood or cell pellets for clinical biomarker studies; Selecting approaches for extending stability based on assays: Immunophenotyping, Functional Assays, Receptor Occupancy (RO); Factors affecting stability: anticoagulant choice, shipping temperature, preservatives Cyto-Chex BCT, Transfix, Smart Tube. 

Topic 3:

Innovations in Mass Cytometry to facilitate scaling to Large Scale Immunophenotyping: Barcoding, Automated Detector Voltage Optimization, Novel Instrument Hardware; Testing new Mass Cytometry for long runs and determining cell loss, marker frequency intensity over multi-day runs; Investigation into MDIPA Panel Stabilization of Whole Blood to determine Sample Stability Limits and Stabilization Blood Collection Tube; Finding balance between sample stabilization and panel compatibility when developing new panels for multiple mouse and tissue studies. 

Topic 4:

Innovative Application of Multiplexed Immunophenotyping by Spectral Flow Cytometry to the assessment of Immunomodulatory impurities; Understanding the principles of assessment of immunomodulatory impurities in Immunogenicity Risk  and existing assays and gaps; Whole Blood and PBMC-based assays  overlook impact on minor but critical cell populations; Innovative method using barcoding to anchor Multiplexed Spectral Flow Cytometry to enable comparative characterization of innate immune response modulating impurities in therapeutics; Proof of concept studies to examine the sensitivity to trace levels of TLR3, TLR7 and TLR9 agonists; Application of multiplexed spectral flow cytometry using mAbs and oligonucleotide drug products.

Topic 5:

Limitation of Traditional NAb assays with binary yes/no result regarding the presence of neutralizing antibodies versus Newer NAb assay Formats; Advanced use CBA like Flow Cytometry in incorporating advanced readouts, providing deeper insights into Clinical Significance of NAbs; Focus on Recently FDA Draft Guidance that emphasizes the need for immunogenicity information Clinically Useful; Unlike assays that simply detect ADA, NAb assays evaluate the host immune response that directly neutralizes the drug's activity.

Topic 6:

Current challenges with NAb CBA Development/Validation due to poor sensitivity, limited drug tolerance, and high variability; Use of  affinity capture elution step to improve drug tolerance of a cell-based cAMP assay; Advance platform method for the detection of NAbs to incretin molecules; Regulatory Requirements to study Neutralizing Antibodies during clinical development; Need for using Cell-based Assays (CBA) for agonist molecules to reflect the mechanistic interactions among the drug, target, and NAb.  enicity data demonstrate the sensitivity and drug tolerance of the NAb assays are appropriate to support clinical studies.

Topic 7:

Focus on major Advantages of Cell-Based NAb Assays versus Limitations of Competitive LBA;Crucial important to overcome current issues in CBA development/validation due to their biological relevance, ability to mimic in vivo conditions; How to improve CBA variability, consistent results, reproducibility, and reduce complexity, matrix effect, drug interference; Is Flow Cytometry a solution to raditional CBA? Competitive LBA issues with physiological relevance, being not able to fully capture the biological complexity of drug interactions; Latest development in Choosing the Right Assay for NAb Testing.

Topic 8:

Innovative application of Receptor occupancy (RO) Assays to Disaggregated Tissue Samples versus traditional RO assays applied to intact human whole blood samples; Need to develop Tissue RO for Drug Target not expressed by the peripheral immune system; Overcoming the lack of literature to help with Method Development; Importance of incubating intact biopsies with saturating amounts of drug to mimic in vivo drug exposure; Tissue RO Assay able to deliver data supporting drug Target Engagement (TE) in relevant tissue in vivo.

Topic 9:

Strategic steps and methods used for assembling an advanced 29-color High Dimensional Spectral Flow Cytometry Regulatory T-cell (Treg) Panel; Development of a specific protocol for Fit-for-purpose (FFP) Validation and application for Clinical Samples analysis; Application of high dimensional analysis incorporating a novel statistical method to determine LLOQ for the identified clusters; Evaluation of the analytical results revealing Novel Regulatory T-cell  Clusters in Normal Healthy Volunteers (NHV) as compared to disease samples.

Topic 10:

Global Harmonization of Spectral Flow Cytometry instruments; Revolutionizing Clinical Cell-Based Assays analyzing more reportables with less blood and by getting harmonious data out of each of instrument for producing accurate quality data; Ensuring accurate data within/between labs by processes/instruments harmonization; Current need to have clinical trials in different sites around the world and issues with samples collected from patients very short shelf life necessitating sample analysis in regional labs.

Topic 11:

Successfully handling/harmonizing Spectral Flow Cytometry & AI/ML-based Analysis as a powerful tool for Receptor Occupancy (RO) assays; Enhanced capabilities to provide more comprehensive and accurate data, for complex modalities, allowing high-parameter (multicolor) analysis, Expanded Panel Design, Deeper Characterization, Improved Resolution and Sensitivity; AI/ML approaches addressing high-dimensional data, Automated Data Analysis, Enhanced Precision and Accuracy Handling Complexity; Considerations & harmonized experience for RO method design, development, and validation, customized for specific cell subsets. 

Topic 12:

State-of-the-art Development/Optimization/Standardization of B Cell ELISpot Assay specifically tailored to detect AChR in patients at the Single-Cell Level; Importance to have the method aligned with current BMV Guidelines & key validation parameters; Implementation of robust normalization protocols and use of high-quality PBMCs, essential lot-to-lot bridging of critical reagents to ensure assay reproducibility for reliable inter-assay comparisons and inter-laboratory harmonization as a reliable immune monitoring tool in clinical trials..

Topic 13:

Progressive optimization of Single-Cell Mass Cytometry workflow for longitudinal studies in the CAR-T Cell Therapy reducing variability and enabling cross-trial comparison; Implementation of Automated Analysis Pipeline and comparison against Manual Gating; Unsupervised analyses comparing manufacturing outcome with phenotype align with findings from automated analysis pipeline; Measuring polyfunctionality via Mass Cytometry in CAR-T Cell Drug Products and creating an analysis pipeline for it; Challenges in identifying unique biomarkers to differentiate Drug Products during the manufacturing process.

Topic 14:

Recent advancements in Flow Cytometry & ELISpot for Single-Cell Analysis focusing on increasing multiplexing capacity, sensitivity, integration with Microfluidic & Artificial Intelligence/Machine learning (AI/ML) to better understand cellular heterogeneity and function; Multiparametric analysis of single cells; ELISpot as gold standard for highly sensitive functional analysis of secreted proteins at the Single-Cell Level detecting low-frequency antigen-specific T cells; Optimizing condition for PBMC sample pre-treatment for highly sensitive, specific, and high-throughput cellular immune response assessment for Gene, Cell & Vaccine Therapies.

2026 White Paper on Cell-Based Assays

Consensus & Conclusions on Cell-Based Assays for 2026 White Paper