Conformational Comparability of Factor IX–Fc Fusion Protein, Factor IX, and Purified Fc Fragment in the Absence and Presence of Calcium
Journal of Pharmaceutical SciencesPublished 23 January 2012
Damian J. Houde, Steven A. Berkowitz
Citations27
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TL;DR
Biophysical results further support the conclusion that fusing an immunoglobulin gamma 1 Fc to rFIX does not significantly alter the higher-order structure of FIX or Fc, Ca binding to FIX, orFc functionality.
Abstract
A long lasting recombinant factor IX -Fc fusion protein (rFIX-Fc) is being developed for the treatment of hemophilia B and is currently in late stage clinical investigation. By limiting injection frequency and maintaining efficacy, rFIX-Fc shows promise as a new therapeutic option for hemophilia B patients. However, before gaining regulatory approval, rFIX-Fc must undergo rigorous analytical and biological testing, in addition to clinical trials. Included in this testing is the need to understand this protein's higher-order structure and dynamics. In this study, we investigated and compared the biophysical properties of rFIX-Fc, rFIX, and Fc using hydrogen/deuterium exchange mass spectrometry and differential scanning calorimetry. Within the limits of these techniques, our results show that structural comparability exists between rFIX and the FIX region of rFIX-Fc. In addition, changes in the structure and dynamics of both proteins, in response to calcium binding, a requirement for FIX function, are also highly comparable. In the case of Fc and Fc region of rFIX-Fc, conformational comparability is also established. These biophysical results further support the conclusion that fusing an immunoglobulin gamma 1 Fc to rFIX does not significantly alter the higher-order structure of FIX or Fc, Ca binding to FIX, or Fc functionality.
Keywords
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