Automated platform buffer screening for multiple proteins on Big Tuna

Importance of the Topic

Buffer conditions critically influence protein stability in biologics development. Traditional manual buffer exchange is laborious and prone to variability, delaying formulation optimization. Automated high throughput screening accelerates identification of optimal buffers, supporting robust development and quality control of new therapeutic proteins.

Objectives and Study Overview

This study demonstrates the use of the Big Tuna automated platform to perform a platform buffer screen for four monoclonal antibodies across six formulation conditions. It aims to streamline buffer exchange, measure exchange efficiency, and concentrate samples, reducing hands-on time while maintaining protein integrity.

Methods and Instrumentation

Big Tuna employs pressure based ultrafiltration and diafiltration with gentle orbital mixing to exchange buffers in Unfilter 96 and Unfilter 24 plates. The protocol alternates filtration, volume measurement via an ultrasonic sensor, and reagent refill until achieving a predefined exchange percentage and final concentration. Protein concentrations were assessed by the Lunatic spectrophotometer using A280 measurements. Stability and aggregation were evaluated by dynamic light scattering on the Uncle platform.

Main Results and Discussion

Big Tuna successfully achieved a 96 target exchange percentage for all samples, with actual exchange ranging from 96.9 to over 99.9 across four mAbs and six buffers in under nine hours. Protein concentrations reached the 5 fold increase target (approximately 50 mg/mL) with high recovery and consistent final volumes (1626 ±115 µL). DLS analysis showed no significant changes in hydrodynamic size or polydispersity after buffer exchange, confirming sample integrity.

Benefits and Practical Applications

• High throughput screening of up to 96 samples with minimal hands on intervention
• Automated cycle time optimization ensures uniform exchange across variable samples
• Integration of concentration and buffer exchange in a single workflow
• Maintained protein stability, supporting reliable formulation development

Future Trends and Opportunities

Advancements may include integration of real time analytical monitoring to further streamline decision making and closed loop control. Expansion of compatible filter formats and miniaturized assays could enable earlier screening of scarce or costly samples. Coupling automated buffer exchange with machine learning for predictive formulation design represents a promising direction for accelerated biologics development.

Conclusion

The Big Tuna platform provides a robust, automated solution for high throughput buffer screening and concentration of antibodies. It significantly reduces manual labor and improves consistency without compromising protein stability, enhancing the efficiency of formulation workflows in biopharmaceutical research.