Related Experiment Videos
Sodium chloride enhances the storage and conformational stability of BDNF and PEG-BDNF
W J Callahan1, L O Narhi, A A Kosky
1Department of Pharmaceutics, Amgen, Inc, Thousand Oaks, California 91320, USA.
Purpose:
BDNF, a noncovalent homodimer, was modified by covalently attaching polyethylene glycol (PEG) with an average molecular weight of 20kDa to the N-terminal methionine. Stability of modified BDNF (PEG-BDNF) in aqueous solution was compared to BDNF after storage at elevated temperature in the presence and absence of NaCl.
Methods:
SDS-PAGE. Light Scattering and Size Exclusion Chromatography were used to assess conformational stability and chemical degradation. In addition, CD spectroscopy was used to follow changes in secondary and tertiary structures upon thermal stress of the protein.
Results:
NaCl containing formulations are more stable than NaCl-free formulations. In NaCl-free formulations, the main degradation product of BDNF and PEG-BDNF had a molecular weight of monomer that was more chemically degraded than the dimer. Additionally, the degradation of PEG-BDNF occurred at an accelerated rate compared to BDNF in NaCl-free environments.
Conclusions:
The addition of NaCl to formulations enhances the shelf-life and conformational stability of both BDNF and PEG-BDNF.