Hydrophobic interaction chromatography (HIC) is widely used to determine drug-to-antibody ratio (DAR), a critical quality attribute of ADCs. However, some ADCs present challenges in DAR determination by HIC due to poor peak separation of DAR variants, with the root causes remaining unclear. We investigated which ADC properties impact peak separation in HIC and identified linker length as one of the structural features significantly affecting the peak separation.

Driving bioanalytical method development is crucial for advancing drug discovery and development. Robust methods ensure accurate measurement of therapeutic candidates, metabolites, and biomarkers in complex biologics. Continuous innovation and optimization drive efficiency, accelerate timelines, and enhance data quality, ultimately enabling safer and more effective therapies.

Structural integrity of an mRNA construct encoding the antigenic protein is key to potency of an mRNA vaccine. Additionally, for vaccines delivered in lipid nanoparticle (LNP) formulations, physicochemical properties of LNPs impact cellular and translation of mRNA to deliver the encoded protein antigen. Measurements of structural and biophysical properties of the mRNA payload and the LNPs can be correlated to protein-expression efficiency (potency) in vitro and immunogenicity in vivo.

Short oligonucleotides are an emerging therapeutic platform for rare diseases, offering high specificity and ease of design. Their chemical modifications impact overall structure, stability, and safety, yet current low-resolution methods provide insufficient assessment of quality attributes. Here, we present NMR techniques for high-resolution structural fingerprinting and compare the results to other analytical methods. We employ univariate and multivariate statistical tools, enabling the detection of subtle variations in their biochemical properties and identifying the sources of these variations. Our work demonstrates NMR as a powerful tool for high resolution fingerprinting of important quality attributes of short oligonucleotide therapeutics.

Developability guidelines primarily reflect conventional antibody architectures, leaving innovative formats with extended CDR loops underexplored. Antibodies bearing long or ultra-long CDR-H3 regions – particularly those forming autonomous antigen-binding domains known as picobodies – offer new therapeutic opportunities but remain poorly characterized from a biophysical standpoint. We examine their developability profiles, defining principles for identifying format-specific liabilities and guide engineering strategies that expand current frameworks for unconventional antibody formats

In the development of AAV-based gene therapies, it is important to obtain a drug product with high purity. CE-Western assays enable increased throughput and automated workflows for the analytical assessment of AAV, such as assays to quantify the relative stoichiometry of viral proteins (VP). Various purified samples from diverse AAV serotypes were analyzed to determine their VP ratio. The ratio of VP3/VP1 in rAAV samples was correlated with biological activity. The results were further supported by LC-MS analyses. We demonstrated that CE-Western can be used as a high-throughput platform to assess the identity, composition, and purity of rAAV drug products.

The rapid development of dual-specific antibody therapeutics is being revolutionized by AI-driven design combined with high-throughput cell binding technologies. AI accelerates candidate prediction, optimizing binding affinity and specificity for dual targets. High-throughput screening enables efficient evaluation of thousands of antibody variants against cell populations. Together, these innovations significantly reduce development timelines and enhance precision.

• Join us for an engaging discussion on emerging trends shaping the future of analytical data.
• Explore how organizations are leveraging advanced analytics to enhance decision-making.
• Learn how AI-driven insights are transforming strategies and enabling smarter, data-driven outcomes.

• ​How effective are current control strategies in managing variability and ensuring quality?
• What emerging tools and technologies are shaping the next phase of characterization control?
•  What does the future of characterization look like? 

Characterizing antibody-receptor interaction kinetics (kon, koff, KD) is crucial for drug discovery, but traditional biophysical methods are not always amenable for complex antibodies or targets. This talk presents alternative cell-based binding assays. We also introduce a novel label-free pre-equilibrium assay to simultaneously determine kon, koff, and KD for up to 30 therapeutic antibodies on live cells using the Gyrolab platform, offering a solution for screening challenging drug formats.

Binding kinetics of therapeutics and its target protein are crucial for the efficacy and safety of the drug. Using surface plasmon resonance (SPR) technology, we performed a competitive SPR chaser assay, a method to study biomolecular interactions with very slow dissociation rate constants (kd < 1E-4 s-1). In this talk, the principle and the experimental setup of the chaser assay will be discussed.

Bioanalytical assays are crucial for therapeutic development, enabling dose regimen determination, efficacy, and safety assessment. Integrating molecular interaction characterization into bioanalytical assay development provides deeper insights into underlying interaction mechanisms, thereby enhancing assay performance, expediting the development process, and ultimately improving therapeutic outcomes. This presentation will highlight case studies that demonstrate the effective application of various molecular interaction characterization tools in supporting the bioanalysis of biotherapeutics, showcasing the benefits of incorporating these tools into bioanalytical workflows.

Residual host cell proteins (HCPs) in biotherapeutic drug products are the process impurities that can compromise stability and safety and must be carefully monitored and evaluated. Current control strategies of HCPs in biotherapeutic drug products involve a combination of upstream process controls, downstream chromatography and filtration, as well as advanced analytical methods such as immunoassays (ELISA) and proteomic mass spectrometry for monitoring and risk mitigation. A risk-based approach, integrating the bioprocessing techniques to remove problematic HCPs and sensitive HCP assays, is essential for ensuring product quality and patient safety.

Host cell proteins are critical attribute for biologics and vaccines. Currently, there are only two methods to analyze this, namely, ELISA for official release method, and mass spectrometry for characterization. However, ELISA method is actually not compatible with anti-sera reagent validation method of 2D western blot. Here we developed quantitative and automated alternative method for HCP, capillary western blot, which is compatible with reagents validation. Assay is compared with ELISA and show that ELISA number is not accurate as compared to capillary western in Vero cell.

I present the CLiC (Convex Lens-induced Confinement) platform for quantitative single-particle and single-cell imaging, combining label-free interferometric scattering (iSCAT) with multi-channel fluorescence. This enables simultaneous measurement of nanoparticle size, mRNA payload, mass, and dynamics in cell-like conditions (Kamanzi et al., ACS Nano 2024, 2021; Boateng et al., Nano Lett. 2025). CLiC supports high-throughput, precision characterization for mechanistic studies and quality control of mRNA-LNP vaccines and therapeutics across manufacturing and biological environments.