A fully integrated Big Kahuna workflow to automate the entire biologics formulation development process

Importance of the topic

Therapeutic proteins are prone to degradation and instability, requiring extensive formulation screening and stability testing. Automated workflows can significantly increase throughput, reduce hands-on time and improve data consistency in biologics development.

Study objectives and overview

This study demonstrates a fully integrated automated workflow for buffer and excipient screening, sample stressing and multi-parameter analysis using bovine serum albumin as a surrogate. A total of 48 formulations spanning five buffer systems (pH 4.5–7.0) and various excipient combinations were prepared in 2 mL serum vials. Samples were analyzed at t0 and after agitation, heat and freeze-thaw stresses.

Methodology

The Big Kahuna system executes automated liquid handling for formulation preparation, aliquoting and dilution. Samples are stressed on the platform under controlled agitation, heating and freeze-thaw cycles. Analytical assays at t0 include pH measurement, viscometry, turbidity, visible particle counting, dynamic light scattering and UV absorbance. Post-stress analyses employ size exclusion and reversed-phase HPLC integrated via the central data management software.

Used instrumentation

• Unchained Labs Big Kahuna automated formulation system with dual robotic workstations
• Lab Execution and Analysis software for workflow control and data linking
• Wyatt DynaPro II for dynamic light scattering
• Molecular Devices SpectraMax microplate reader for UV/Vis concentration measurements
• Agilent 1100 series HPLC for size exclusion and reversed-phase chromatography

Main results and discussion

Initial turbidity values ranged from 5 to 13 NTU, viscosities near 1.2 cP and particle counts of 0–4. RP-HPLC confirmed high purity of BSA in all formulations and SEC-HPLC showed minimal aggregation at t0. Heat stress induced the most pronounced increases in polydispersity, turbidity (>100 NTU), visible particles and browning, particularly in formulations at low pH (4.5–5.0) and with sorbitol/NaCl. Agitation and freeze-thaw produced modest changes. Overall protein stability remained high, indicating formulation components primarily governed observed stability differences.

Benefits and practical applications

Automating the entire workflow reduced hands-on scientist time by approximately 4- to 6-fold compared to manual methods, while standardizing sample handling and ensuring data integrity through centralized tracking. The high-throughput system accelerates formulation optimization, supports consistent data reporting and frees researchers to focus on experimental design and interpretation.

Future trends and potential applications

Further integration of orthogonal analytical methods, machine learning-driven formulation design, cloud-based data analytics and remote laboratory operation are anticipated to enhance automation capabilities. Extending similar workflows to a broader range of biologics, peptides and complex modalities will streamline development pipelines and enable adaptive formulation strategies.

Conclusion

The fully integrated Big Kahuna automated workflow provides a robust platform for high-throughput biologics formulation development. By combining automated compounding, stressing and multi-modal analytics within a single system, laboratories can achieve reproducible results, significant time savings and improved data confidence without additional staffing resources.