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.

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.

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.