This talk presents a case study in the practical application of automation and mechanistic chromatography modeling in early-stage (FIH) purification process development. High-throughput screening and modeling are used in conjuction to support PD and subsequent tech transfer of a cation-exchange polishing step for an mAb with high aggregate burden. Importantly, the model is shown to be useful in adapting operating conditions to changing feed impurity levels due to cell-line improvements, thereby improving process outcomes at scale. Overall, the proposed workflow allowed for timeline acceleration, process improvement, and de-risked tech transfer, enabling the toxicology study to support IND.

Mechanistic and data-driven approaches are reshaping biologics development, yet process characterization (PC) remains experiment-intensive. An in silico PC workflow is presented based on mechanistic chromatography models for automated, scale-aware characterization of downstream unit operations. Applied to Protein A capture and cation exchange polishing, the framework combines targeted experiments, model calibration and validation, surrogate modeling, Monte Carlo sampling, defect-rate analysis, global sensitivity analysis, and equivalence testing against manufacturing-scale data. The workflow determined proven acceptable ranges and critical process parameter classification entirely in silico, with close agreement to experimental PC. The study supports in silico PC as an enabler for Bioprocessing 4.0.

Adequate mixing and buffer exchange during ultrafiltration/diafiltration (UF/DF) are critical for robust downstream bioprocess performance, particularly when mechanical agitation is constrained by equipment or operating limits. In this study, computational fluid dynamics (CFD) modeling was applied to evaluate hydrodynamics, mixing efficiency, and buffer exchange performance in a large-scale UF/DF vessel operated without active agitation during diafiltration.

3D-printed chromatography columns provide advantages over conventional packed bed resins, in particular, control over the multiscale structure that can be tailored to the needs of the separation. However, possible disparities between design and fabricated columns lessens this benefit. We apply 3D imaging to evaluate and compare printed structures to original CAD files to refine the design process, creating more effective separation capabilities for emergent biological products.

In vitro transcription enables large-scale mRNA production but generates difficult-to-remove double-stranded RNA (dsRNA) impurities. Existing chromatographic approaches for dsRNA removal are limited by scalability and manufacturing cost. We report a mixed-mode PrimaS monolith chromatography method that selectively clears dsRNA from 1–10 kb mRNA under RNA-compatible conditions, preserving integrity and supporting its potential as a platform purification solution.

This presentation introduces Hydrophobic Interaction Chromatography (HIC) as a powerful, aqueous alternative to traditional anion exchange and reverse-phase methods for ASO purification. We demonstrate how retaining the 5' hydrophobic blocking group enables a high dynamic binding capacity, allowing for an efficient bind-and-elute process. Our data highlights a systematic optimization of salt types and stepwise gradients to achieve >90% product yield with superior resolution of failure sequences and process-related impurities.

This talk shares an enhanced pre-use sanitization strategy for AVB Sepharose resin in adeno-associated virus (AAV) production. By evaluating various alcohol, acid, and NaOH combinations against environmental microbial contaminants and assessing the identified combination in AAV resin re-use studies, we offer a solution that enhances microbial control without compromising resin performance or product quality.

This dataset evaluates how extended affinity resin binding times impact adeno-associated viral (AAV) vectors. We assessed key quality attributes, including yield, aggregation, post-translational modifications, and transduction efficiency. While serotype and ligand strongly influence certain outcomes, host-cell impurity clearance trends consistently across multiple AAV clades. Ultimately, these findings deepen process understanding to optimize downstream AAV manufacturing.

We have developed a scalable, enzyme-free, closed workflow using fibrous chromatographic media before tangential flow filtration (TFF) to selectively remove extracellular chromatin (DNA/histones) from extracellular vesicle (EV) preparations. This strategy prevents proinflammatory responses while preserving EV yield and identity. Moreover, the use of fibrous chromatographic media improves purity, safety, and manufacturability, de-risking late-stage development and enabling GMP-compliant, large-scale EV production.

Hexameric IgGs are a class of complex biologics with unique pharmacokinetic and pharmacodynamic properties arising from their high valency (up to 12 antigen-binding sites), large molecular weight (~900 kDa), and increased hydrodynamic radius. Despite their therapeutic promise for certain indications, technical challenges in purification and analytical characterization continue to hinder the broader application of hexameric IgGs. This talk aims to demystify these challenges by providing a comprehensive analysis of hexameric IgG molecules, accompanied by illustrative examples and case studies in which key technical issues were identified and successfully resolved. It will offer practical insights and serve as an introduction for researchers interested in the discovery and development of large, complex biologics.

We present the evaluation and optimization of MabSelect SuRe 70 for capture purification of a bispecific antibody in comparison with an alternative Protein A resin. MabSelect SuRe 70 demonstrated higher dynamic binding capacity, improved specific product recovery, and increased monomer purity. Further process improvements were achieved through optimization of elution conditions to enhance resolution and host cell protein (HCP) clearance.

This presentation discusses trispecific antibody (tsAb) intensified and/or continuous purification process design, development, and manufacturing. Challenges associated with molecule complexities and product-related impurity are addressed. Process design and development strategy and achievement are addressed. A prototype of tsAb continuous manufacturing is proposed to enhance process efficiency, productivity, and reduce cost of goods. The ultimate goal is to produce adequate amount of biologic drug substance and product with high purity, quality, safety, and potency for clinical and commercial needs.