The study explains how Gryolab Protein Technologies helped Sanofi's Biologics Development group achieve the increased efficiency they sought to integrate into their downstream process development workflows.

To overcome regulatory burdens associated with host cell proteins during early downstream process development of candidate monoclonal antibodies, one Sanofi researcher discussed how the company worked with Gryolab assays to implement a completely revamped high-throughput analysis workflow.

The problem with host cell proteins

Host cell proteins are process-related impurities generated during the production of monoclonal antibodies and are a critical quality attribute for selection of monoclonal antibodies during bioprocess development. During process purification, the majority of host cell proteins are removed; however, residual amounts can remain in the product, particularly for monoclonal antibodies. Impurities caused by host cell proteins can cause risks for product quality, safety, and efficiency. As such they remain a critical regulatory point for health authorities.

The enzyme-linked immunosorbent assay (ELISA) is the gold standard approach to monitoring host cell proteins during bioprocessing. It is a well-established and robust analytical method that is highly sensitive.

However, ELISA offers limited pathways for automation, with significant hands-on time and analysis time with multiple incubation and washing steps. A high-throughput assay that provides a similar dynamic range as ELISA could be beneficial to downstream bioprocess development workflows.

The case study explains how a team of researchers from Sanofi's Biologics Development group, led by Suzana Petrovic, have worked to change process development by increasing the use of automated approaches and minimizing host cell proteins during early developmental stages. With the challenges that many bioprocessing teams face for multiproject development under reduced timelines, the Sanofi team turned to Gyrolab technology to address these concerns.

Turning to high-throughput technology

Gryolab Protein Technologies was formed in 2000 and offers an immunoassay system for nanoliter-scale protein quantification. Streptavidin beads are used to build the immunoassay with a microfluidic design that contains 15 nanoliter affinity bead columns within a microstructure housed on CDs. Flow-through technology uses centrifugal force and capillary action to deliver a sample/reagent that is detected with laser-induced fluorescence. The company offers a high-throughput system that can be implemented to accelerate process development for its customers.

A head-to-head comparison of Gyrolab's assays versus ELISA reveals that the platform can significantly increase throughput with reduced hands-on-time while achieving an increased dynamic range. This would ultimately translate to a reduced need for replicate samples and lower volume requirements, according to Petrovic.

But Petrovic didn't just take Gryolab's word for it. Her lab conducted benchmark studies where they tested actual molecules under development for host cell protein identification using Gryolab's CHO-HCP E3G kit versus its standard in-house ELISA Cygnus F550 assay. They found that Gryolab technology was able to achieve similar results to ELISA, but with high-throughput efficiency.

Based on these data, Sanofi implemented the Gyrolab assay at all three Global Biologics Development locations. After implementation, they were pleased with the highly comparable results they observed across different equipment at all three sites.

Improving other areas of the workflow

The technology allowed the team to provide full high-throughput support analysis for early downstream process development. However, after they integrated the system into their workflow, they observed a bottleneck where manual sample preparation was no longer compatible with the increased-throughput system. Therefore, the team transitioned to the Agilent Bravo liquid handling system, which automates dilution for 96 samples in just 10 minutes. This allowed the team to eliminate manual dilution for high-throughput host cell protein analysis and massively reduce retesting due to pipetting mistakes. With this change, they increased analytical capabilities by a factor of five, without losing quality.

A second bottleneck that Petrovic's team identified after implementation of the Gryolab assay was data treatment, which became more complex with an increased number of samples and preparations. The team was previously using an older version of Gryolab software that only provided partial data analysis and required significant operation time.

However, once they upgraded to a newer version of Gyrolab software these issues were resolved, and they can now quickly perform full data analysis for host cell proteins. This version of the software (v 8.1.3) includes a data manager and sample mapping for ease of analysis, said Rob Durham, PhD, Gyrolab's Protein Technologies director of service and scientific support.

Overall, the Sanofi Biologics Development team has fully implemented high-throughput support for early downstream process development with a quick turnover rate of just one day. These changes will allow them to reduce the time needed to introduce new molecules to clinical studies.

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