Protein Folding, Binding and Lyophilization by Real-time PX2+ Fluorescence Sensing for Biopharmaceutical GMP Applications
Fluorescence spectroscopy has been used to assess changes in the tertiary structure of proteins in the solution and solid state.1 The sensitivity of fluorescence to the protein tryptophan environment has made it a useful tool for studying protein conformation, binding or complexation and stability.1-3 While fluorescence spectroscopy has been used to assess such changes, single response at a single excitation-emission wavelength combination via a LED based photometer, such as our PX2+ equipped with an in-line front surface probe, affords in situ monitoring of various protein phenomena. That is, the lower cost and sustainable PX2+ with a front surface probe or a single use flow cell enables real-time monitoring of protein folding, lyophilization and associated stability with GMP manufacturing unit operations or within a quality or development laboratory environment.
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Ramachander, R.; Jiang, Y.; Li, C.; Eris, T.; Young, M.; Dimitrova, M.; Narhi, L., Solid state fluorescence of lyophilized proteins. Analytical Biochemistry 2008, 376 (2), 173-182.
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Sharma, V. K.; Kalonia, D. S., Steady-State Tryptophan Fluorescence Spectroscopy Study to Probe Tertiary Structure of Proteins in Solid Powders. Journal of Pharmaceutical Sciences 2003, 92 (4), 890-899.
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Dufour, C.; Dangles, O., Flavonoid–serum albumin complexation: determination of binding constants and binding sites by fluorescence spectroscopy. Biochimica et Biophysica Acta (BBA) – General Subjects 2005, 1721 (1), 164-173.