In addition to high-throughput and multi-product analytical methods, the use of robotics for sample preparation automation may further reduce sample analysis time and cost. There are several companies that provide liquid handling automation instruments, including LEAP Technologies and TECAN. Figure 6 shows a LEAP Technologies CTC PAL liquid handling system, which is intended for HPLC and GC usage and capable of fraction collection and injection. The CTC PAL instrument is capable of on-the-fly sample preparation, such as protein dilution and digestion. On-the-fly sample preparations are viable if the sample preparation takes less time than the analytical method. For sample preparations that take longer than the analytical run time, batch sample preparation can be performed using robotic liquid handling systems such as the TECAN Freedom EVO, which can handle multi-well plates for increased sample throughput. Robotic liquid samplers can increase reproducibility, efficiency, and safety compared to manual handling of samples.
The final steps to most characterization workflows include data analysis and report generation. Several software packages are available that are designed to reduce the time necessary to complete post-data acquisition tasks. For liquid chromatography applications, commercially available chromatography data software, such as Dionex's Chromeleon chromatography management software and Waters Corporation's Empower chromatography data software, include features such as automated peak integration and one-click report generation. The Chromeleon software also has an extension pack that includes validation report templates and sequences, enabling increased automation of method validation. More companies are implementing electronic laboratory notebooks, which have advantages over traditional laboratory notebooks such as ease of data sharing and collaboration, streamlined review and witnessing processes, standardized documentation, and long-term data preservation.
SUMMARY
MAbs are under development for a wide range of indications, and ever-growing biotechnology pipelines have greatly increased demand for high throughput, multi-product protein characterization methods. By taking a holistic approach to improving the protein characterization process, one can identify strategies to increase value and reduce the development time of an analytical method. Implementing high-throughput and multi-product liquid chromatography methods that use available knowledge and resources typical in a biopharmaceutical facility can address the increasing demand for protein characterization studies, and these methods can be readily validated following regulatory guidelines. Further technological advancements in sample preparation throughput and data handling automation adds value to the overall characterization process, which will enable faster product development and shorter timelines to clinic.
ACKNOWLEDGEMENTS
We would like to acknowledge the many people involved in our work, including but not limited to Ed Bouvier at Waters Corporation, Mark van Gils and Guillaume Tremintin at Dionex Corporation, and Peter Smith at LEAP Technologies, for technical input and assistance.
Jennifer C. Rea, PhD, is an associate scientist, G. Tony Moreno is a senior research associate, Yun Lou is a research associate, Rahul Parikh is a research associate, and Dell Farnan, PhD, is a senior scientist and associate director, all at Genentech, Inc., Protein Analytical Chemistry, South San Francisco, CA, 650.225.6378 , [email protected]