Stability & formulation screening

The problem

Misbehaving proteins can unfold, aggregate and ruin everything, but the right protein construct will hold it together and save the day. Once you’ve got the right protein, picking formulations is the next big hurdle. Screening for the best proteins and formulations requires having all the right data to advance the best candidates.

Outdated ways of looking at protein stability take way too much sample and time, and often deliver only part of the story. To keep an eye on all the ways that protein stability can go wrong, unfolding and aggregation have to be measured at the same time on the same sample. Plus, the earlier you can weed out bad candidates, the better – so it’s critical to use tiny amounts of protein.

The right tool for the job

Uncle is the all-in-one stability platform that enables more than twelve different applications to quickly study protein stability - on only 9 µL of sample. Fluorescence, static light scattering (SLS) and dynamic light scattering (DLS) modes all read the same sample and combine to give powerful insights such as thermal melting, aggregation, and sizing.

To get the whole story on protein stability you need to combine SLS and fluorescence. While fluorescence sees unfolding, SLS will spot aggregation the instant protein molecular weight changes. Run up to 48 samples at a time and get results in less than 2 hours so you can screen more candidates, formulations, or conditions to quickly figure out which is the most stable.

The proof

How Uncle spots stability problems

A DSF experiment on Uncle starts with using DLS to look at size and size distribution right off the bat, so you know if your protein is good to go or if it’s already suffering from aggregation. Next, as Uncle heats up your samples, fluorescence picks up when protein unfolding occurs and calls out any melting temperatures (Tm). SLS then shows when aggregation begins as an aggregation temperature (Tagg) – and that can sync up with Tm1, Tm2, or happen independently from any Tm. Because SLS intensity is related to molecular weight, it's incredibly sensitive to aggregation and is the best way to see exactly when aggregation begins. After the thermal ramp maxes out temp, DLS can be used once more to quantify how bad aggregation has gotten.

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Mix it up

Uncle allows you to compare proteins, formulations, and everything else that impacts stability. A simple amino acid mutation or change of buffer can increase or decrease melting temperatures or can change unfolding behavior so that a protein with two melting temperatures now has three. Similar kinds of changes can happen with sugars, salts, surfactants, pH shifts or other protein hazards like freeze/thaw cycles.

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Stack the deck

Beyond the classic DSF experiment for protein stability, the flexibility of three detection technologies teamed up with temperature control makes Uncle a jam-packed toolbox of protein stability. Unfolding and aggregation data is just the gateway to isothermal stability, protein self-interaction, refolding, viscosity, and more. Uncle has over a dozen applications designed to rapidly extract the most possible data from the smallest sample volume of protein.

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Uncle

Cracking stability using a pile of one-trick, protein-hungry tools is a ton of work. Uncle combines 3 different measurement modes — fluorescence, Static Light Scattering (SLS) and Dynamic Light Scattering (DLS). So you can crank out all your data in just a few hours, and use way less protein. All the info you’ll get makes picking the best formulation, protein, or viral vector a piece of cake.

Want more info?

Want to learn more about how Uncle makes it easy to screen for the most stable protein or best buffer?