Single cell omics is a powerful technology for the understanding of biological systems. In this talk, studies from our laboratory focused on capturing chromatin accessibility, gene expression ,and protein abundance from monoclonal antibody CHO cell lines will be presented. Single cell omics platforms and computational approaches for data analysis will also be discussed.

Lipid metabolism plays a key role in cellular processes that are central to achieving high recombinant protein titres. Processes such as secretion, cell division, and endoplasmic reticulum size and function are highly dependent on lipids and their metabolism. In this approach, we have used genetic engineering techniques to optimise lipid metabolism in CHOK1-SV cells which has been successful in expanding the endoplasmic reticulum and ultimately enhancing recombinant protein titres between 1.5 and 9-fold.

Macromolecular complexes are cornerstones of most, if not all, biological processes in cells. We will illustrate how the CrispR/Cas9 editing technology can be used for gene tagging in order to introduce affinity tags and facilitate the purification of proteins/macromolecular assemblies expressed in physiological conditions. We will also discuss tagging proteins with fluorescent reporters in view of imaging and functional proteomics applications.

There is a need for a better and more cost-effective method for rAAV production to meet the high demand in the gene therapy field. We address this issue by creating an inducible rAAV producer HEK293 cell line that does not need further transient transfection with plasmid DNA or infection with a helper virus.

Virus-like particles (VLPs) have proven potent vaccines. However, their complex self-assembly can make their development challenging. Here we describe progress in the use of E. coli cell-free synthesis platforms to expedite these activities for this product class. Two examples will be given: picornavirus-like particle vaccine candidates stabilized by N-myristoylation and universal influenza vaccine candidates based on tandem-core hepatitis B core antigen VLPs.

The presentation will highlight the components of the cell systems employed for generations of research cell lines and CHO manufacturing cell lines, and recent developments aiming at enhancing the quality and titer of produced complex biotherapeutic proteins.

Cell line development (CLD) represents a central part of a biologics development process. Hence, accelerating the CLD process would have substantial impact on overall timelines. Integration of the semi-targeted integration (STI) technology into our CLD platform enabled manufacturing of a therapeutic antibody at 2,000L scale in less than 3 months. Next, by using a clonally derived production cell line, a process with ~5g/L was implemented in less than 9 months.

Chinese hamster ovary cells are commonly used for therapeutic proteins production. Major challenge of cell line development is to obtain stable high-producing clones. Therefore, we set out to develop new CHO host cell line that would be selected based on the growth characteristics as well as productivity and secretion potential. To assess cell productivity and secretion capabilities, we used genetic reporter that allowed us for high-throughput screen of CHO cells.

In this presentation, it will be shown how machine learning can exploit the dynamic information on metabolomic and process data to support the selection of highly productive cell lines during bioprocess development and scale-up. In this way, high productive cell lines and the biomarkers that are most related to cell productivity are identified already at the early stages of experimentation.

In-licensing of new molecules may come along with unforeseen CMC challenges. In the case study at hand, we observed significant product fragmentation using a producer cell line from an in-licensing partner. To ensure clinical supply and gain a scientific understanding, potential root causes and solutions were investigated, including the fast generation of a new producer cell line. The exploitation of transposase-mediated semi-targeted transgene integration facilitated the identification of stable producer pools free of fragmentation in a competitive project timeline. Thus, fast cell line development can tackle CMC difficulties, without stretching project timelines.

Biopharmaceutical and AAV production in CHO/HEK293 challenges their translational, secretional, and metabolic boundaries. We show the importance of ER-mitochondria signaling to fulfill energy demand of secreted products. Further, we exemplify how systems and synthetic biology can be used to define limiting genes and tune their expression improving activity of therapeutic enzymes 100-fold by OMICS and engineered TIS and regulatory elements for precise expression.

mRNA based products represent a novel emerging modality with the potential to treat the previously untreatable. This talk will outline and present case data on the development of a flexible and scalable manufacturing process for mRNA-LNP products. This will include the development of the analytical toolkit to support process development and manufacture, which encompasses biochemical, structural and biological activity assays for product characterisation.