Secretory IgA (SIgA) plays an essential role in the defense against pathogenic invasion in human. Production of SIgA for therapeutic purpose can be a novel way to tackle threats brought by viruses and bacteria. We have developed a fast and robust method to produce functional SIgA in CHO-cells with comparable titers to IgGs. We demonstrated efficient antigen neutralisation for these SIgAs. By targeting Hot Spot in vitro, we generated BAC-based expression vectors, which integrated in multiple copies into the CHO genome devoid of detrimental host chromatin effects. This BESTcell platform allows development of stable high-yield production cell-lines within 3 weeks.

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By combining leap-in transposase mediated gene transfer with a well-proven host cell line, we have successfully established the Rentschler Expression Platform. Based on case studies, we will demonstrate how we leverage this promising technology for a fast and efficient screening platform. The enhanced gene expression due to customized vector design, inherent stability and optimal project timelines through seamless integration into our well-characterized USP platform process, translates to optimal therapeutic protein production.

Membrane proteins are important drug targets (GPCRs, ion channels), yet are notoriously difficult to work with. We have developed a novel one-step approach for the incorporation of membrane proteins directly from crude cell membranes into lipid Salipro particles. This direct approach, termed Salipro DirectMX, presents new opportunities for de novo development and characterization of biologics and small molecules, including phage display, B cell sorting and cryoEM.

Ebola GP1/2 and the Spike protein of SARS-COV-2 are typical virus surface antigens. Current protein production technologies for such subunit protein antigens are insufficient to meet the unprecedented global demand for critical ingredients. Using optimized CHO-based technologies, we developed a GMP-compliant process for production of such complex proteins. In weeks-timeframes we were able to produce grams quantities of fully active Ebola GP1/2 and SARS-COV-2 Spike protein trimers. The Ebola GP1/2 trimer generated sterilizing protection in vivo against Ebola infection; the spike trimer induced high antibody titers against SARS-COV-2 in vivo and showed extreme sensitivity and specificity in diagnostic tests.

Intranasal vaccination is considered as promising way to induce mucosal & systemic immunity against SARS-CoV-2 since it resembles the natural route of infections. Different protein variants including a polypeptide fusion to target immune cells in the mucosa were produced. The latter increased the effectivity of immunization 10-fold in combination with the toll-like receptor agonist Riboxxim as adjuvant. The new platform XPOVAX allows the generation of safe, potent and easy-to-administer vaccines

Recently, baculovirus-free expression systems have been employed for more simple and fast protein production in High Five insect cells. This plasmid-based expression results in high protein yields particularly in regard of secreted target proteins and challenging target proteins like trimeric Spike protein of SARS-CoV-2. Several optimization steps to increase the performance of the expression system will be presented highlighting potential applications.

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Serology is a powerful tool for monitoring outbreaks of pandemic diseases like COVID-19.  Serology assays can identify the number of asymptomatic patients and track the progress of the pandemic and vaccination campaigns.  The key to COVID-19 serology assays is high-quality protein reagents including the SARS-CoV-2 spike, RBD, and nucleocapsid.  We highlight production and optimization of these critical reagents to produce higher quality antigens and more sensitive assays for detection of antibodies.

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In order to produce Ichnos’s monospecific and BEAT (Bispecific Engagement by Antibodies based on the T-cell receptor) antibodies, 2 and 3 different chains have to be co-expressed, respectively. Moreover, Discovery activities as well as developability assessment require substantial amounts of purified candidate molecules that are challenging to obtain with transient production. As a result, we used the Leap-in transposase technology to generate stable pools while fine-tuning the expression of each chain to ensure proper molecule assembly and obtain titers in the g/L range. To do so, different transposon designs, promoter and leader peptide strengths were tested and their impact on titers and product quality was assessed.

Lead strain selection is a critical step during microbial process development, initially based on product titer because product quality evaluations are challenged by the timely availability of sufficiently purified material. The capability of a well-proven E. coli expression platform has been enhanced through the use of a scale down/high throughput approach to protein purification using robotics, to enable strain selection based on parallel evaluation of product titer and quality.

Although monoclonal antibodies are considered as standard molecules in biotherapeutic industry, some of those drugs are still difficult-to-express molecules. Here, we are highlighting the development of a difficult-to-express monoclonal antibody and its challenges for CHO cell line generation. Therefore, protein engineering and modulation of vector architecture are presented as approaches to improve product yields. Moreover, a new cell line development concept in CHO going beyond random transgene integration is highlighted to further boost the expression of a difficult-to-express molecule.

Continuous processing is used as a tool to maximize productivity in many industrial sectors. Regarding microbial continuous cultivations, subpopulation formation causes instabilities in long-term cultivations. Cascaded continuous cultivation was found to facilitate stable protein expression using microbial hosts, however mechanistic knowledge of this cultivation strategy is scarce. In this presentation we demonstrate a method workflow to reduce workload and accelerate the establishment of stable continuous processes with E. coli BL21(DE3).

This presentation provides a straight forward approach for the development and manufacture of  market oriented products in the field of biotherapeutics, including mammalian and microbial technologies with specific aim on which are the main items to be fulfilled in a development program.