J.KNIBBE,1 D.WEINFURTNER,2 J.JAEHRLING,2 R.OSTENDORP,2 B.BROCKS,2 W. FRIEß,3 M.WIGGENHORN1
1Coriolis PharmaService GmbH, Am Klopferspitz 19, 82152 Martinsried, Germany
2MorphoSys AG, Lena-Christ-Str. 48, 82152 Martinsried, Germany
3Ludwigs-Maximilians-Universität München, Butenandtstr. 5, 81377 Munich, Germany
Posted with permission from Michael Wiggenhorn,
Purpose:
The aim was to evaluate the use of high throughput based formulation (HTF) development for a liquid monoclonal antibody (Mab) formulation in comparison to a traditional vial-based formulation approach. Furthermore, a particle measurement technology using Micro-Flow Imaging™ was integrated.
Methods:
Liquid formulations of Mab (differing in pH and excipients) were prepared in matrix vials using an automated pipeting workstation. A Design of Experiments (DOE) approach was performed using the SAS JMP 8 software and a central composite design. All accelerated stress tests (elevated temperature, freeze-thawing cycles and mechanical stress tests by agitation) were performed in a 96-vial matrix plate and in 10R vials. HP-SEC, DLS, turbidity, and UV measurements were performed in 96-well plate formats.
Results:
Our results emphasized the necessity to include multiple stress methods in the formulation screening. Different stress-specific degradation pathways require to be addressed differently by modifying the formulations, i.e. with respect to added excipients. The high number of samples to be analyzed by HP-SEC, UV, fluorescence and DLS could be managed straightforward by applying micro plate formats and liquid handling robotics. A major challenge remained the accurate quantification of subvisible particles was crucial to identify the most stable formulations. Overall, the new formulation approach resulted in about 40% reduction in the protein material required compared to the classical vial based methods and allowing testing a much wider range of formulations.
Conclusions:
Multiple stress tests are required to develop a robust protein formulation and that a combination of a wide range of analytical techniques and stress conditions is required to assess the impact of the different constituents on formulation stability. The new formulation strategy resulted in comparable outcomes as the classical vial-based approach in identifying stable formulations for the antibody. MFI proved to be valuable to assess subvisible particles. Overall, relative simple modifications in the formulation development process, including high throughput based assays allowed for a significant reduction in time and material requirements.
PRESENTED AT:
2010 AAPS National Biotechnology Conference, San Francisco CA, USA
2010 AAPS NBC_CoriolisPharma_Orthogonal stress methods and particle measurement for HTP.pdf
2010 AAPS NBC CoriolisPharma Orthogonal