Aunty protein stability characterization tool

Significance of the topic

Protein stability assessment is essential in the development and quality control of biotherapeutics, vaccines and research reagents. High throughput and multiparameter analysis accelerate formulation screening and candidate selection while conserving precious samples.

Objectives and study overview

This study demonstrates the performance of the Aunty platform in comprehensive protein stability characterization. It evaluates thermal melting, aggregation, colloidal stability, long term monitoring and viral vector analysis in a single automated workflow.

Methodology

Samples are loaded into a 96-well quartz plate using 8 µL per well. A thermal ramp from 15 to 95 °C is applied at defined rates while intrinsic or reporter dye fluorescence is recorded at excitation wavelengths of 280 and 470 nm. In parallel, static light scattering (660 nm laser) and dynamic light scattering monitor aggregation onset, size distributions and hydrodynamic diameters. Colloidal stability metrics kD, B22 and G22 are extracted from concentration series during isothermal or temperature ramp experiments. Long term stability protocols record signals over hours or days to detect slow unfolding or aggregation. Fluorescence assays track genome release and capsid integrity in viral vector samples.

Instrumentation

The Aunty instrument integrates LED excitation sources (280 nm UV and 470 nm blue), a full range spectrometer (250–750 nm), a 660 nm laser and avalanche photodiode detectors. Temperature control is accurate to ±0.1 °C with a range of 15–95 °C and heating rates from 0.1 to 10 °C/min. Automated plate handling supports reading a full 96-well plate in one minute with <2% CV for melting temperatures.

Main results and discussion

Aunty delivers high resolution melting temperature (Tm and Tonset) and aggregation (Tagg and Tsize) profiles with simultaneous fluorescence and light scattering data. Colloidal stability assessments differentiate protein formulation behavior via kD, B22 and G22. Long term stability tests reveal formulation dependent trends over extended periods. Viral vector assays accurately detect payload ejection temperatures and capsid unfolding events. Data throughput and sensitivity enable rapid identification of optimal candidates.

Benefits and practical applications

• Ultra high throughput screening of formulation conditions and variants
• Minimal sample consumption (8 µL) and robust chemical compatibility
• Integrated multispectral fluorescence and light scattering modalities
• Real time monitoring of thermal and isothermal stability
• Quantitative colloidal stability and aggregation metrics
• Specialized assays for viral vector integrity

Future trends and potential applications

Integration with automated sample preparation systems and data analytics platforms will further streamline screening workflows. Machine learning algorithms may predict stability profiles from multiparameter datasets. Expanding applications to membrane proteins, nanoparticles and complex biologics will broaden the impact of this technology.

Conclusion

The Aunty platform offers a comprehensive, high throughput solution for protein stability characterization, combining fluorescence and light scattering in a single instrument. Its sensitivity, speed and low sample requirements support accelerated biotherapeutic development and quality control.

References

No external references were provided in the source document.