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СпутникиРНSpaceXРоскосмосNASAHigh-throughput protein stability measurements with Aunty Blog High-throughput protein stability measurements with Aunty A protein stability measurement is the biophysical characterisation of how well a protein formulation resists melting and aggregation under thermal, chemical, or other stresses; this measurement is a critical element of developability testing, which identifies the most stable protein candidates and formulations…
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A protein stability measurement is the biophysical characterisation of how well a protein formulation resists melting and aggregation under thermal, chemical, or other stresses; this measurement is a critical element of developability testing, which identifies the most stable protein candidates and formulations to advance through drug development. High-throughput stability platforms make it possible to run protein stability measurements across dozens or hundreds of conditions simultaneously with minimal sample consumption.
When it comes to protein stability measurements, speed is king, but only if it comes with fidelity. Traditional stability methods have drawbacks: some are slow, don’t give the entire picture, or use too much sample. Aunty blasts through those issues to crank out actionable results at breakneck speeds and claim the crown as queen of protein stability.
Aunty’s full spectrum DSF gives you the choice between intrinsic or dye-based methods to determine a protein’s melting temperatures. Static and dynamic light scattering (SLS & DLS) run in parallel to each other, and with fluorescence, to spot aggregates at any point of an experiment. The three methods work together with Aunty’s one-of-a-kind quartz 96-well plate to give you the full protein stability profile of all the samples every minute in an SBS-format plate from only 8 μL of each sample.
Heating up proteins in solution causes them to unfold, exposing hydrophobic amino acids to an aqueous environment. This transition changes their fluorescence behavior – often shifting the peak to higher wavelengths. You can track the shift to find a melting temperature (Tm) and use it to rank the conformational stability of formulations, constructs, mutants, or even conjugates.
Protein aggregation often happens alongside unfolding. As proteins aggregate, the SLS intensity of the solution will increase and the average molecular diffusion will decrease. DLS tracks that decrease in diffusion to determine the hydrodynamic size of particles and the distribution of particle sizes. Parallel monitoring of unfolding and aggregation is the key to protein stability measurements on Aunty.
Formulation stability studies are often about finding the right balance of excipients. Arginine in a formulation reduces protein aggregation and improves colloidal stability but also drives protein unfolding. Aunty lets you assess how much of an impact those different effects have on your antibody. Adding arginine to bovine IgG (blue) decreased the Tm, compared to no arginine (green), but the SLS intensity showed an increase in the temperature of the onset of aggregation, Tagg. Side-by-side DLS measurements showed a later size increase temperature, Tsize, in presence of arginine. Same-time protein unfolding and aggregation on Aunty means you won’t miss a thing and can draw the right conclusions regarding stability of mutations, modifications, conjugations, or formulations.
Screen confidently with remarkable reproducibility
Maximizing throughput while minimizing sample consumption are major goals of protein stability measurements, but so are high levels of accuracy and precision. Aunty plates are made of quartz and have impeccable optical performance and chemical compatibility with all common biologics reagents. Sealing the plates after loading is easy and protects your samples from evaporation and contamination. This entire process, from loading samples to sealing plates to running experiments, is automation-ready. Reproducibility is no sweat with Aunty; its Tms have CVs ≤1% and it heats with temperature accuracy ±0.1 °C.
Identify stable biologics or formulations in a snap
When you use Aunty’s Analysis software to graph Tm versus Tagg, you can immediately tell which samples have good conformational stability, colloidal stability, or both. You can also see the spread between the replicates of each sample, giving you confidence that you’re making the right call.
In an experiment with 7 combinations of antibody and excipient, a trastuzumab biosimilar without excipients (purple) had the highest Tm and Tagg while a pembrolizumab biosimilar (gray) had the lowest. Every biologic will have its own formulation preference and Aunty’s all-quartz 96-well plate makes it the fastest, highest throughput tool to find the sweet spot.
Conclusion
Aunty’s combination of full-spectrum DSF, high-sensitivity SLS and DLS, and precise thermal control makes it the most efficient platform for high-throughput protein stability measurement available today. Aunty’s 96-well plate maximizes your throughput while minimizing sample consumption to just 8 μL per well. Thermal ramp behaviors of your sample are highly reproducible, with Tm CVs ≤1%. By combining Tm, Tagg, and Tsize, you'll churn through any number of replicates and quickly recognize the samples with optimal stability profiles to accelerate developability and formulation screening, helping you bring novel, life-saving therapies to the clinic even faster.