Advantages of using FIDA when oligomerization might be an issue
Know the oligomeric state of the biomolecule or particle you are working with in 3 minutes
Many proteins are composed of multiple subunits and thus oligomerization is rather the norm than the exception. The association between these subunits can vary in strength and duration.
In some cases, oligomerization dependents on environmental conditions, such as concentration, temperature, and pH; other proteins oligomerize dynamically in response to a stimulus, such as a change in protein or nucleotide binding, hydrolysis or post-translational modifications such as phosphorylation and glycosylation. Protein oligomerization is a key parameter for a number of reasons, including functional control, such as allosteric regulation, stability and the establishment of higher-order complexity.
When working with oligomeric proteins, it is essential to know the actual state of the proteins that you are analyzing. Current technologies do not allow for direct confirmation of protein oligomeric state and the analytical steps required in many current technologies may end up affecting the oligomeric state.
Because the Fida 1 is based on direct, in-solution detection of the size of proteins and complexes which binds to the ligand that uniquely identifies your target, you will always get quantitative data confirming the oligomeric state of the proteins you are analyzing as part of your functional characterization, quantification etc. of oligomeric proteins. Get more information about the FIDA technology here.
Quantitative confirmation of oligomeric state
With the Fida 1’s detection of the absolute size and ability to screen the concentration dependence on oligomerization you always know the oligomeric state of the protein, you areanalyzing.
Oligomeric analysis directly in complex solutions
Oligomerisation may be triggered by the complex environment, or you are manufacturing proteins with a specific oligomeric state. With the Fida 1 you can analyse directly in plasma, serum, cell lysate, fermentation media etc.
Temperature imposed change of oligomeric state
The Fida 1 allows you to control the temperature from 5 degrees to 44 degrees with a tolerance of 0.1° C. Thereby it is easy to study how temperature changes might impact the oligomerisation
Examples of Fidabio data generated in automated workflows
FIG 2: TNFa-alexa488 (trimer 3.2 nm) binding to adalimumab in 10 and 20 % v/v human plasma in phosphate buffer, 0.067 M, pH 7.4 with 0.1 % HAS, Kd = 1.0 – 1.7 nM, complex size: 8.7-9.7 nm
FIG 3: The figure shows the change in hydrodynamic radius (nm) as a function of buffer pH and therefore, the concentration of hydrogen ions (H+). The change in hydrodynamic radius indicates a pH dependent transition of a monomeric state of the protein to an oligomer. Single measurements were performed. R2 = 0.993, Rh, monomer = 1.08 nm, Rh, oligomer = 3.19 nm.
Let us discuss your work with oligomeric proteins
