Biopharmaceutical product development requires a real-time storage period of at least 18 to 24 months in order to demonstrate its long-term stability. In the interest of reducing time and costs corresponding to such long-term storage experiments, there is a strong need to find short-term protein parameters that generate similar information on long-term physical protein stability. One of these short-term protein parameters that shows great promise is the protein cloud temperature (Tcloud). Tcloud is a known representative of protein-protein interactions, which play a dominant role in long-term physical protein stability. In addition to a shorter time frame to generate the desired stability information, material use and experimental effort should be minimized to obtain an efficient screening method. This can be achieved with a high-throughput and low volume approach. An experimental setup to determine Tcloud that incorporates all these desired characteristics is currently not available. The presented work will show an experimental method to detect Tcloud in a low volume and high-throughput manner. This was achieved by combining an automated imaging system and a cryogenic device that is compatible with 96-well microtiter plates, leading to a sample volume of 24 μL. Image-based data evaluation was automated, which resulted in a full plate evaluation time in the order of magnitude of minutes. The performance of the experimental and analytical approach was evaluated via a robustness study and the obtained parameter to represent Tcloud was validated. Furthermore, the work presents a case study employing the developed method to investigate long-term physical protein stability in a shorter time frame. The combined results of this work display the applicability of the low volume high-throughput Tcloud detection method as part of a short-term analytical screening toolbox that optimizes the search for long-term stable protein formulations, thereby advancing short-term strategies for efficient formulation development.

Apparent protein cloud point temperature determination using a low volume high-throughput cryogenic device in combination with automated imaging

Macromolecular Chemistry: The Second Century

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ACS Spring 2020 National Meeting & Expo

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