An automated methodology for assessment of protein stability using nanograms of protein

In this talk, we demonstrate how the Fida 1 platform can be utilized for probing conformational stability of therapeutic proteins using as little as 40 ng protein per measurement combined with walkaway automation. We report C[m]-values as well as the free energy of unfolding for adalimumab, rituximab, and human serum albumin at different pH-values. The developed method measures the change in hydrodynamic radius and intrinsic fluorescence of a protein during in-line denaturation with guanidinium chloride.

The conformational stability of proteins plays a key role in the understanding of living systems as well as in the development of therapeutic proteins. Protein stability data is typically obtained by observing relative changes in optical properties such as fluorescence and/or scattering under thermal or chemical perturbations. The Fida 1 instrument utilizes absolute measurements of hydrodynamic radius and intrinsic fluorescence applying Taylor dispersion analysis (TDA) and LED-UV fluorescence detection. The Fida technology is a “first principles” methodology meaning that assay readout can be predicted from the protein structure (e.g., from PDB files). The simple workflow and low sample consumption (40 ng protein) of the Fida methodology makes it ideal for assessing protein characteristics related to stability in early drug development or when having scarce amounts of sample.