To do this, a stream of your fluorescently labelled protein is introduced alongside an auxiliary stream
These streams flow in parallel and because there is no convective mixing the only way your protein can migrate into the auxiliary stream is by diffusion, the rate of which will depend on the size of the protein. Small proteins will diffuse very rapidly, and large proteins and aggregates more slowly.
At the end the streams are re-split, and at this point the degree of diffusion is fixed. The quantity of protein in each stream is then determined by the fluorescence from the label. The ratio of the fluorescence between the two streams gives us the protein's hydrodynamic radius (Rh)
Fluidity One-W can measure proteins in buffer and in crude solutions like cell lysates or biological fluids, because only the labelled species is detected.
If we repeat the test using a mixture of labelled protein and unlabelled binding partner we can observe the degree of binding due to the change in average size. Only species including the labelled protein are detected and measured.
Titrating the binding partner against the labelled protein gives a binding curve and automatically generates a KD value. The hydrodynamic radius for the unbound protein and protein complex are also calculated.
By measuring your proteins and their interacting partners in a close-to-native, biological background, you can expect more transferable results that accurately represent natural biological processes.
Work in crude biological mixtures with minimal preparation, while controlling for off-target binding. Gain valuable insights at the earliest possible stage.
Gain meaningful insights into protein complexes and their formation with less sample and in less time.
Solution state measurement coupled with high sensitivity means you can study proteins that are problematic for other systems such as:
-Multi protein complexes
-Intrinsically disordered proteins
Assess the quality of your protein complex even in early stage fractions, allowing you to optimize protocols early. Use insights on protein size, state and yield to rationalize how you move forward before you begin time consuming purification.
The minimal sample prep, easy-to-use design and wide dynamic range means you will spend less time setting up, and more time generating the data you need.
The Fluidity One-W is currently in development, and we’re enrolling early access partners to help us define the specification of the instrument based on their applications. If you'd like to learn more and potentially participate then get in touch
*The Fluidity One-W is currently in development. Exact specifications are not yet confirmed and may be subject to change