Membrane proteins such as GPCRs pose a challenge for in vitro study due to their hydrophobic nature and dependence on lipid environments. For these proteins, nanodiscs can be generated as a substitute reagent, but their quality drives ultimate experimental success. Adimab scientists systematically compared nanodiscs produced in-house with commercially available products to understand variability in reagent quality. Using a combination of dynamic light scattering (DLS), biolayer interferometry (BLI), and polymer screening, we developed an optimized workflow that delivers uniform, functional nanodiscs ideally suited for GPCR analysis and antibody discovery.

Key insights and outcomes:

• Commercial nanodiscs displayed inconsistent particle sizes and aggregation levels, as revealed by DLS, underscoring the need for greater control over nanodisc assembly.
• DLS analysis guided purification decisions by identifying early elution fractions (~20 nm radius) as the most uniform, while later fractions showed unacceptable heterogeneity.
• In-house nanodiscs maintained antigen functionality with strong binding to target antibodies and no non-specific interactions.
• Adimab’s in-house nanodiscs achieved higher yields and superior homogeneity, with preparations incorporating Amphipol 18 emerging as the most effective polymer.
• Our integrated workflow combines polymer selection, DLS-based quality assessment, and functional validation to ensure reproducible nanodisc performance.

By directly addressing the variability observed in commercial preparations, Adimab’s optimized in-house system establishes a reliable, analytics-driven framework for producing stable, functional nanodiscs. This approach not only improves reproducibility and assay fidelity for membrane protein studies but also enhances the robustness of antibody discovery campaigns targeting complex transmembrane antigens.

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