Development of a Venofer AUC Analysis protocol and comparison between Venofer and Biosimilar
Challenge
The study aimed to develop an Analytical
Ultracentrifugation (AUC) protocol to compare commercial Venofer with a Biosimilar candidate in
alignment with the FDA's guidelines on Iron Sucrose.
Challenges faced included the high sucrose
concentration in the buffer which can affect sedimentation, the high analyte concentration
leading to non-ideality, light scattering affecting optical signals, and rotor speeds
affecting diffusion and sedimentation balances.
Solution
A rotor speed of 13,000 rpm was adopted, which minimized
the effects of sucrose gradient formation and achieved a balance between diffusion and
sedimentation.
Wavelength optimization was conducted, with 300 nm
selected for optimal observation, reducing light scattering while maintaining good Xenon
lamp efficiency.
Different dilutions were tested, and a range of 1:2000
to 1:6000 was found optimal, addressing the issues of concentration-dependent non-ideality.
The effects of aging and sonication on the
samples were also examined to ensure the validity of the results.
Conclusion
A robust AUC protocol was successfully developed to
compare Venofer with the Biosimilar candidate.
By addressing the inherent challenges of high sucrose
and analyte concentrations, light scattering, and rotor speeds, the study ensures accurate,
reproducible results that align with FDA guidelines.
Future comparisons should utilize this
protocol for consistent results.
