To improve understanding and increase options in developing a successful production cell line with desired product quality profile, it is important to develop diversified CHO host lineages with differences in cell growth and protein production in responding to medium and process conditions. In addition, developing predictive OMICS tools and integrating into quality control strategy are highly useful in selecting highly productive recombinant cell lines with the desired protein quality profile.

Genetic code expansion holds the promise to revolutionize the life science and biomedicine through expanding macromolecular chemical diversity outside the natural space. We developed an engineered Escherichia coli cell-free system which allows rapid sequestration of selected native tRNA isoacceptors and subsequent reassignment of the liberated sense codons to unnatural amino acids. This represents a powerful tool for numerous practical applications including production of constrained peptides, antibody-drug conjugates and novel enzymes.

Cell-free expression systems became key tools for the production of membrane proteins and other challenging targets. We have developed protocols to insert membrane proteins already co-translationally into supplied nanomembranes of defined composition. The generated protein/nanoparticles are suitable for biochemical and structural studies and contacts with detergents or other artificial environments are avoided. The strategy is exemplified with membrane-inserted phage toxins currently being explored as potential inhibitors of bacterial cell-wall formation.

A novel nanoparticle can encapsulate, fold and release proteins of a wide range of sizes, charges, and disulfide content to produce improved yields and improved specific activity. Our index manuscript (Nature Communications | 8: 1442) described the concept and we have a much larger manuscript under review now which we fully expect to be published this year.

TipSort technology (an advanced, in-house robot pipetting algorithm) optimizes pipetting throughout a dynamic 96-position plate or tip tray. In-house CAD design married to 3D-printing fuels the crafting of custom lab inventions, spawning innovation and saving dollars. A fully-loaded Hamilton Vantage Robot enhanced with custom configuration/integrations and a built-in machine learning driven scheduler will support the future of small-scale protein expression analysis in the laboratory.

ProBioGen's travel equipment for the shortest journey to even higher mountains of cell productivity (with a high elevation base camp at pool stage)

In this talk we will assess consecutive advancements of the transposon system: genome targeting, activity and template recognition and show how this optimized system combined with the a refined ALS CellCelector platform affects the cell line development workflow and its outcome for antibodies, proteins and viral vectors.

With most top blockbuster drugs therapeutics being glycoproteins, there is a growing interest in engineering their glycan structures for improved safety, efficacy, and manufacturing. Using our systems biology approaches, we can predict the modifications needed to effectively glycoengineer proteins. We further have explored the more global impacts glycoengineering has on the host cell, thus helping to define the design space of CHO-produced glycoproteins.

A novel public resource that presents all codon usage, codon-pair usage and human tissue specific codon usage and codon-pair usage will be discussed. Examples of investigation areas which could greatly benefit from this resource will be provided, such as biotherapeutic development, tissue-specific genetic engineering and genetic disease prediction.

There are no therapies that reverse the proteotoxic misfolding events that underpin neurodegenerative diseases including amyotrophic lateral sclerosis (ALS) and Parkinson’s disease (PD). Hsp104, a hexameric AAA+ protein from yeast, solubilizes disordered aggregates and amyloid but has only limited activity against human neurodegenerative disease proteins. Here I will describe approaches we have employed to re-engineer Hsp104. The engineered variants can dissolve aggregates, restore protein localization, suppress proteotoxicity, and attenuate dopaminergic neurodegeneration in animal models.

Conventional mammalian protein expression strategies using transient expression or random genome integration with gene of interest, followed by high-throughput colony screening and expansion, are laborious and/or time-consuming. To address those issues, we have developed a series of synthetic biology toolkits to transform mammalian protein expression system engineering. Our powerful platform technologies are versatile and can be adapted to different mammalian cell types and biomanufacturing settings to optimize complex therapeutic protein production.

A mutation in a ribosomal protein that we discovered in cancer cells enhances the levels of ribosomal protein translation and its fidelity, and reduces cellular proteasome activity in lymphoid cell models. To validate these findings in mammalian cell lines that are commonly used in the recombinant protein production industry, we engineered the ribosomal mutation into CHO and HEK293 cells. A 69-155% increase in production of 3 recombinant proteins was confirmed in HEK293 cells.