AUTOMATED SAMPLE PREPARATION AND DATA HANDLING
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.
Figure 6. LEAP Technologies CTC PAL sample preparation robotics system. The system interfaced with a Dionex U3000 HPLC, with
both CTC PAL system and HPLC controlled by Dionex Chromeleon chromatography management software. MAb samples were digested
and diluted by the CTC PAL and injected directly onto the HPLC for IEC analysis.
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.
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.
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
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,
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