NEHA N. PARDESHI,1 JOHN F. CARPENTER1
1Department of Pharmaceutical Sciences, University of Colorado, Denver
Posted with permission from Neha Pardeshi,
Purpose:
From previous studies we know that pumping with a positive displacement filling pump causes particle formation of therapeutic IgG products. The particle formation is due to the heterogeneous nucleation caused by the steel nano-particles shed from the pump. In the current study we tested the hypothesis that pumping-induced particles would increase the sensitivity of the formulation to post-pumping agitation stress.
Methods:
We used intravenous immunoglobulin G as a model therapeutic protein product. A positive displacement piston pump was used for the filling operation, and an orbital shaker at an RPM of 350 was used for the agitation studies. Particle sizes and numbers were obtained with micro-flow imaging (MFI), and size exclusion chromatography (SEC) was used to quantify levels of monomer and soluble aggregates.
Results:
Agitation in the absence of pumping induced particle formation. However, a much higher number of particles were observed with agitation of the pumped sample. There was a time dependant increase in particle number and size during agitation. The pumped and agitated sample became visibly cloudy after 7 days. After 15 days, there were more than 2.5 million particles of 1 micron and larger size. Initially, although there was substantial number of particles formed, a loss of protein could not be detected by SEC. However, at latter time points, the particles agglomerated and were of sufficient mass that detectable loss of monomer due to “insoluble aggregates” was measured with SEC. Again, the magnitude of this damage was much greater in the pumped then in the unpumped samples. No soluble aggregates were detected.
Conclusions:
Our results support the hypothesis that particles forming during filling pump operation can increase the sensitivity of therapeutic protein formulations to post-pumping stress.
PRESENTED AT:
2010 AAPS National Biotechnology Conference, San Francisco CA, USA
2010 AAPS NBC_UofC_Effect of mechanical stress on therapeutic proteins during fill-finish ops.pdf
2010 AAPS NBC UofC Effect of mechanical stress on