Upstream process development in a high throughput automated bioreactor system

CHO cell based high expression platform

The generation of a stable, robust and high producing CHO cell line is the start of CMC drug development. Boehringer Ingelheim’s established high expression (BI-HEX®) technology platform provides outstanding quality combined with high expression. Our BI-HEX® platform leads to a fast-track delivery of the research cell bank (RCB) about eleven weeks after transfection with the toxicology material supply starting five months after transfection using a clone that has been tested for stability, biosafety and BI-HEX® platform fit.

The BI-HEX® platform has an extensive track record of over ninety cell line development projects, with a significant number leveraging the efficiency of the new CHO-K1-based GS-knockout host cell “BI-HEX® CHO-K1GS.” This cell line is our current gold standard used to support the development of more complex proteins such as bispecific antibody formats and non-mAbs (e.g. enzymes, growth- factors) in addition to standard monoclonal antibodies.

Cell line development process

Cell line development process

BI-HEX® CHO-K1GS host cell line
History and background of the well-characterized, reliable and high producing BI-HEX® CHO-K1GS host cell line.

In-silico analysis service
It all starts with the amino acid sequence: the in-silico prediction of CMC bottlenecks
The option of in-silico amino acid sequence analysis to potentially boost expression and identify possible bottlenecks in CMC drug development can be offered. Boehringer Ingelheim has also demonstrated that protein stability as well as further CMC properties can be enhanced with in-silico guided sequence optimization. The single amino acid exchange(s) are based on the analysis of the amino acid interactions in the predicted protein structure. A further benefit is that these exchanges often result in titer increases as e.g. folding of the protein is improved. The analysis of the protein structure itself already provides our formulation and downstream experts valuable insight in the expected CMC properties. This knowledge can be used to assist during development and to define the ideal process setting.
[ref. Baur 2020 DOI: 10.1080/19420862.2020.1787121]

Advanced expression vector
Expression vector for genetic engineering
The BI-HEX® platform consists of an expression system with Boehringer Ingelheim’s proprietary GS platform expression vector. This expression vector uses well-elucidated transcriptional enhancers and comprises new technologies such as microRNAs for expression of difficult-to-express proteins.
[ref. Fischer et al. 2017]

Superior cell culture medium
Boehringer Ingelheim’s proprietary chemically defined media and feeds are animal-component-free and used during cell line generation, scale-up and transfer to production facilities. The media and feeds are designed to meet the demands of the cell during cloning, expansion and production. Several years of experience with scale-up and transfers to production facilities, an extensive characterization of media, feeds and supplements as well as qualification of suppliers provide high reliability and in-depth knowledge of the media and feeds.

High throughput automation
Boehringer Ingelheim offers a state-of-the-art laboratory equipped with automatic plate handlers, cell sorters, imaging platforms, cell counters and liquid handlers. This allows us to minimize handling errors and increase overall efficiency during cell line generation.

Monoclonality assurance
Boehringer Ingelheim’s cell line generation platform ensures a high probability of monoclonality of the production cell by two orthogonal methods resulting in a probability of monoclonality of > 99.9 %. The method was successfully filed and approved for several projects.
[ref. Fieder et al. 2017]

Sophisticated platform fit and high producer screening
Boehringer Ingelheim’s automated high throughput screening for growth, titer, product quality as well as fit to the BI-HEX® platform allows for the identification of highly productive and optimal performing clones within about eleven weeks. Screening selection criteria can be customized to the needs of the individual product.

Cell line stability assurance
During cell line development also the stability of the production clones is assessed. The stability is confirmed by a comprehensive stability study confirming genetic and phenotypic stability. The results may be reported in clinical and commercial filing application.

Early material supply
Downstream production and analytical development are important steps in the development process and require early representative material. Boehringer Ingelheim offers this starting material with the selection and scale-up of the top performing cell pools. The material is used to shorten timelines, define the process for tox material and GMP clinical production and provide necessary methods for material release.

Filing strategy
Supporting our customer’s regulatory filings is an essential part of Boehringer Ingelheim´s collaboration with its customers. A large number of clinical and commercial applications based on the BI-HEX® platform have been accepted by local and global health authorities.

Readiness for clinical supply or process readiness
Boehringer Ingelheim’s top clone selection workflow provides for delivery of a final clone with platform fit to allow fast tox material production and process transfer to the GMP facility for clinical material manufacturing. Moreover, the cell line generation platform has the ability to select a clone viable for commercial product manufacturing up to 12,000 liters.