A humanized monoclonal antibody fragment (Fab) formulation becomes turbid and contains excessive sub-visible particles upon reconstitution with purified water after being subjected to shear stress prior to formulation, filtration, and lyophilization.  Filtration with a sterile 0.2 �m filter prior to lyophilization did not reduce the turbidity of the reconstituted solution to levels present in the unstressed controls.  This indicates that sub-micron particles smaller than 0.2 �m could be passing through the filter membrane and contributing to the solution turbidity after reconstitution.  The size and quantity of sub-visible particles were characterized via light obscuration, flow microscopy, and dynamic light scattering.  Additionally, reconstitution of the lyophilisates with 0.1% w/v polysorbate 20 significantly reduced the solution turbidity, indicating that hydrophobic interactions could be contributing to the formation of sub-visible particles.  The use of other surfactants and stabilizers in the formulation and diluent and their effect on final solution turbidity was also studied in a multivariate (DOE) approach to identify main effects and further elucidate the mechanism of sub-visible particle formation.

Investigating sub-visible and sub-micron particles formed during shear stress,      filtration, and lyophilization of a monoclonal antibody fragment

247th National Meeting (2014)

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.