Emerging novel biotherapeutics, which are typically difficult-to-express, require improvements for high-yield production. In this study, we conducted a genome-wide fluorescence-activated cell sorting (FACS)-based CRISPR knockout screening in bispecific antibody (bsAb)-producing Chinese hamster ovary (CHO) cells. The screening identified ATF7IP and SETDB1 genes, which are binding partners for H3K9me3-mediated transcriptional repression. Knockout of the ATF7IP-SETDB1 complex enhanced bsAb productivity by 2.7-fold and monoclonal Ab productivity by 3.9-fold without affecting product quality.

This study explores the effects of protein glycation on immunotherapy for glioblastoma multiforme (GBM). We investigate how glycation, a common post-translational modification, influences immune cell function, tumor microenvironment, and treatment response. Our findings provide insights into the potential role of glycation in cancer immunotherapy and may inform future therapeutic strategies.

Cytokine/antibody fusion proteins (termed immunocytokines) assemble intramolecularly to bias cytokine signaling behavior through multi-layered structural and molecular effects, and overcome common issues of free cytokines when used as therapeutics. Immunocytokines require special considerations with respect to their production to avoid oligomerization and/or aggregation. This modular approach, based on interleukin-2 (IL-2), can be extended to any cytokine of interest for a broad range of biomedical applications.

• Construct design and misfolding prevention
• Leveraging machine learning and data optimization
• Cell-free systems for complex protein synthesis

Thermothelomyces heterothallica C1 is a well-known industrial enzyme production host able to reach 120 g/l enzyme levels in a 6-7-day process. We have developed the C1 technology suitable for low-cost manufacturing of therapeutic and vaccine proteins including virus-like particles (VLPs), nanoparticles, and individual antigens. Production levels ranging from several hundreds of mg/l to 20 g/l of secreted protein have been obtained.

Fusion proteins (MBP, GST, etc.) are frequently used in recombinant protein production pipelines to improve soluble yields of target proteins. The removal of fusion proteins is typically accomplished enzymatically with site-specific proteases (TEV, 3C, etc.). While this can be highly efficient, removal of these enzymes post-fusion removal can complicate target protein purification. Enzyme immobilization represents one potential solution to this problem. Here we present strategies for the incorporation of immobilized enzymes into recombinant protein production workflows.

We have developed a screening method to identify reactive cysteines in recombinant proteins that is coupled to a high-throughput protein expression and purification platform. Proteins purified in 96-well plates are pre-screened against representative covalent fragments to detect the presence of reactive cysteines. Proteins identified with reactive cysteines are prioritized for scaled-up reagent production and more comprehensive screening, optimizing both reagent utilization and screening resources.

Recent pandemics have accelerated vaccine development, highlighting recombinant Virus-Like Particles (VLPs) for strong, durable immune responses and dose sparing. Scalable high-yield VLP production reduces costs, crucial for global vaccine manufacturing. ExpreS2ion Biotechnologies developed a VLP-based COVID-19 vaccine, which was later validated in Phase III clinical trials, and is now applying VLP technology to HER2 positive breast cancer, demonstrating its broad applicability. We continue to enhance our platform by modifying glycosylation pathways in S2 cells to significantly modulate the product's immunogenicity.

In-cell NMR spectroscopy is a potential method to investigate the behavior of therapeutic proteins and their targets at atomic resolution in living cells. We used in-cell NMR combined with site-specific 19F-labeling, enabled by cell-free protein synthesis, to explore the membrane-associated states of the Ras protein in living cells. This approach allowed us to characterize the conformational states of Ras depending on its nucleotide-bound states and oncogenic mutations.

Protein-biochemistry-related support of chimeric antigen receptor (CAR) T cell therapy programs from early development through commercialization requires effective project management, nimble business practices, and excellent cross-functional communication. This is facilitated by several tools, including a laboratory information management system (LIMS), SMART goal setting practices, and an environment that properly balances individual and teamwork-oriented tasks.

With the growing realization of clinically effective multispecific therapies, there has been a steady rise in the exploration of complex biologics specifically in immunology and oncology. This poses a unique challenge for drug development, with double or triple the number of antibody discovery campaigns needed per project, followed by the subsequent evaluation of libraries of multispecific antibodies with diverse formats, target clone properties, geometries, and valencies. In this study, we will showcase how robust functional assays using primary cells can be miniaturized and run in high throughput to provide early insights into what makes a potent multispecific drug.

Protein engineering is a highly iterative process, with multiple rounds of hypothesis-driven experimentation leading to better hypotheses in subsequent rounds on an overall trajectory toward a fitness optimum. Our Self-Driving Autonomous Machines for Protein Landscape Exploration (SAMPLE) platform automates the hypothesis, experiment, and data interpretation steps in a closed, autonomous loop, enabling researchers to focus on the overall experimental design rather than the lengthy iteration process, accelerating progress.