We will describe the development of an end-to-end automated solution for high-throughput intact mass spectrometry analysis of monospecific biologics, utilizing a combination of external analysis software, internal bioinformatics-enabled tools, and an enterprise-wide data registration platform. The process improves QC timelines from 2 weeks to 1 day, eliminates over 80% of scientist’s hands-on time, enables AI/ML training through consistent formatting and uploading, and allows for analysis of 2,000+ molecules per week.

Production of recombinant proteins involves maintaining their integrity through purification and downstream applications. This is especially challenging for autolysis-prone peptidases. We developed an activity-guided purification strategy that suppresses autolysis, thus enabling production of intact KLK7, a serine peptidase, without resorting to catalytic mutations or inhibitors. Further investigation helped identify novel biological substrates of KLK7 and inhibitory antibodies, demonstrating a robust approach for purifying labile recombinant enzymes.

Biologic manufacturing processes and formulations include a series of purification, filtration, and buffer exchanges. Excipients from therapeutic protein formulation serve critical roles in stabilization, cryoprotection, and osmolality. We describe a simple and rapid quantitative 1H benchtop NMR method for measurement of excipients without the need for complex sample preparation. The method is relatively quick which lends itself to being Process Analytical Technology (PAT), with at-line measurements or in-process QC testing.

Small GTPases require purification in defined nucleotide-bound states for structural and biochemical studies, yet conventional workflows separate purification and nucleotide exchange, increasing time and sample handling. We developed a tandem-affinity chromatography method that integrates nucleotide loading during purification, producing high-purity K-Ras4B and H-Ras proteins suitable for downstream applications. This presentation will highlight the workflow, implementation considerations, and its extension to additional GTPases and nucleotide-binding proteins.

Infrared Matrix-Assisted Laser Desorption Electrospray Ionization (IR-MALDESI) has recently been established as a platform for ultrahigh-throughput MS analysis. Herein, we report the applications of an IR-MALDESI system in early biologics discovery workflow. It accelerates screening and reduces bottlenecks compared with traditional LC-MS workflows. Its ability to analyze thousands of samples quickly makes it a powerful tool for improving decision-making, boosting efficiency, and advancing biologics discovery at scale.

Predicting antibody behavior in cation exchange chromatography (CIEX) remains challenging using conventional metrics like theoretical pI. We utilized high throughput analytical CIEX data from hundreds of antibody binding domains along with protein language model embeddings and sequence- and structure-derived descriptors to develop predictive ML models. These models provide improved tools for building blocks selection during multispecific antibody design and enable increased purification throughput by providing better prediction of CIEX separation.

In biologic discovery, purification strategies must be tailored to varying scales and business needs, especially when considering higher throughput workflows. This talk outlines core chromatography principles, compares batch versus column approaches, and evaluates tip-based, pre-packed, and other innovative techniques needed to automate these processes. Emphasizing time, cost, binding/elution efficiency, and ease automation, the study helps make fit-for-purpose choices that close industry gaps and strengthen end-to-end biologic discovery workflows.

A common challenge in discovery is the expression of new target proteins in the form of insoluble inclusion bodies, where refolding is necessary for recovery of the active target. This work describes a strategy for combining refolding with affinity capture, purification, and self-removal of the affinity tag in a single, streamlined process. A case study on a difficult target that requires insoluble expression in E. coli will be presented.

This presentation explores strategies for purifying complex protein therapeutics beyond traditional Protein A approaches, addressing challenges from early screening through process development and scale-up. It will highlight efficient, scalable purification workflows for challenging molecules, including alternative capture and polishing strategies, platform approaches, and process optimization. Attendees will gain practical insights into selecting purification strategies that improve recovery, purity, scalability, and overall development efficiency.

A case for the use of 384-well scalability and identifying key parameters for process monitoring in high-throughput formulation development. Establishing the best ways to inform on formulation stability and potency monitoring strategy for high-branched studies.

Machine learning and generative AI are transforming antibody discovery by enabling prediction of vast antibody sequence spaces, increasing demand for scalable experimental validation. AstraZeneca has reengineered its automated infrastructure to support high-throughput generation of transfection-ready constructs. This presentation will describe the strategic development and deployment of AstraZeneca’s end-to-end automation platforms to reduce cycle times, expand library generation, and enhance production and delivery capacity for simple to complex antibody formats, enabling validation of data-driven predictions at discovery scale.

Producing diverse therapeutic proteins efficiently requires more than a standardized workflow. This presentation will explore how reusable approaches across construct design, expression, purification, and characterization can be combined with molecule-specific decision-making. Drawing on experience leading protein science teams in therapeutic development, I will discuss how to recognize when platform methods are sufficient, when adaptation is needed, and how knowledge sharing across programs reduces repeated work. Attendees will gain practical perspectives on balancing standardization with scientific judgment to accelerate problem-solving and improve development efficiency.

Production of protein complexes and/or their constituents can be challenging given that the solubility and stability of the complex components may be dependent on their physical association with each other. Cellular localization (e.g. extracellular, membrane-anchored) and unique structural features (e.g. disulfide bonds, post-translationalmodifications) are additional complicating factors. AlphaFold-based structure prediction was used to guide construct design and prioritize candidate domains and putative subassemblies of a large extracellular microbial complex for experimental study. Conditions for large-scale expression and purification were determined by a 96-well expression/solubility screen.

Protein production workflows should be designed around the downstream question the reagent must answer. This presentation will explore how expression and purification choices influence protein quality, functionality, and suitability for structural, biochemical, and biophysical studies. Emphasis will be placed on defining fit-for-purpose requirements early and using characterization to confirm that produced proteins are suitable for their intended discovery applications.