We developed a bacterial strategy for the expression and purification of a SARS-CoV-2 spike protein receptor binding domain (RBD). Bacterial cytoplasm is reductive and this is problematic when the recombinant protein of interest requires complicated folding and/or processing. The CyDisCo system bypasses this issue by pre-expressing a sulfhydryl oxidase and a disulfide isomerase, allowing the correct folding with disulfide bonds for protein integrity and functionality. We show that it is possible to quickly and inexpensively produce an active RBD in bacteria that is capable of recognizing and binding to the ACE2 receptor as well as antibodies in COVID-19 patient sera.

Rapid and comprehensive strain selection and an early process development program utilizing the Pelican Expression Technology™ were key to establishing the foundation for late-stage success for Rylaze™, a recombinant Erwinia chrysanthemi asparaginase for the treatment of acute lymphoblastic leukemia or lymphoblastic lymphoma in patients who have developed hypersensitivity to E. coli-derived asparaginase. Pelican Expression Technology™, a Pseudomonas fluorescens-based protein expression system is a robust and scalable platform for recombinant protein production.

Cost-effective methods of expression and purification of SARS-CoV-2 spike and its fragments that preserve antigenic properties are essential for development of COVID-19 serology tests. We developed a method of purification of S319-640 fragment containing RBD expressed in E. coli from the inclusion bodies. The antigenic properties of the resulting product are similar to those of the RBD-containing fragment expressed in human cells.

RNAs have emerged as a novel class of medications with unique mechanisms of actions. Current RNA research and development are restricted to the use of chemo-engineered RNA mimics made in vitro, which are different from natural RNAs produced and folded in vivo. In this talk I will present our novel RNA bioengineering technology for high-yield, large-scale and cost-effective in vivo fermentation production of true biologic RNA agents carrying payload RNAi molecules.

We have generated and characterized CSL040, a recombinant soluble and truncated form of human Complement Receptor 1 (CR1) that potently inhibits all three pathways of the complement system and represents a new and attractive therapeutic candidate for complement-mediated disorders. CSL040 attenuates disease development in multiple animal models, and we demonstrate that sialylation-dependent pharmacokinetics and differential complement pathway inhibition are hallmarks of CSL040 activity in vivo.

Recent advances in synthetic biology and artificial intelligence promise to dramatically impact the way that pharmaceuticals are developed.  In this presentation I will discuss how Absci is leveraging the power of these technologies to address key pain points in both how protein-based biologics are discovered as well as how cell lines for manufacturing are designed.  

We developed a novel platform (TOBI) centered upon an inert tissue factor scaffold for production of heteromeric and multi-functional fusion protein complexes for therapeutic uses. We will present our characterization of HCW9218 (a bifunctional TGF-ß trap/IL-15 protein complex) and HCW9201 (a fusion complex combining IL-2, IL-15, and IL-18), created based on the TOBI platform, to demonstrate the versatility and potential utilities of these fusion proteins as immunotherapeutic against cancer.

CHO cell volumetric productivity has significantly increased in the past two decades. However, turning these cells into high-producing factories has created an overall metabolic burden. In response to transgene expression pressure, CHO cells undergo phenotypic and genetic changes for continuous improvement.  Selexis’ SUREtechnology Platform™ provides a solution for overcoming the challenges of high productivity tied to metabolic adaptability. 

We discuss the challenges in high-throughput protein production for small and large molecule drug discovery. We demonstrate the parameters and design space required to generate high-quality proteins for HTS, antibody discovery, in vivo and developability studies. Supported by our industry-leading platforms, the Protein Sciences department at WuXi Biologics provides production services utilizing various expression systems for the generation of monoclonal, bispecific and multispecific antibodies, and other recombinant proteins.

Each protein secreted by a cell depends on up to hundreds of different host cell proteins for its synthesis, folding, post-translational modification, and transport. However, this secretory pathway machinery required can differ considerably. Using computational modeling and omics data integration, we have developed a pipeline to identify the host machinery that can be augmented to improve secretion of diverse recombinant biotherapeutics of interest. 

The ability to identify and characterize therapeutic antibodies targeting multi-pass membrane proteins is hampered by their often-difficult expression and purification.  Historically membrane proteins have been extracted from cell membranes using detergents, however the presence of detergent can hamper many downstream applications. We examined novel methods for membrane protein stabilization, including SMALPs, nanodiscs and amphipols, to isolate a model multi-pass membrane protein and evaluated their utility in lead antibody identification.

Bispecific antibodies are an emerging class of biologics that have shown great promise in cancer treatment. Yet often during preclinical development, many lead molecules fail due to poor biophysical/biochemical properties. Here a phage-display method has been developed, in which engineering of the cyno cross-reactivity, stability and expression of a bispecific molecule was tackled simultaneously, generating multiple lead molecules that showed significant improvement in all three properties.

Chinese Hamster Ovary (CHO) cells are one of the major workhorses for Transient Gene Expression (TGE) of recombinant antibodies. Through a matrix evaluation of multiple factors including inoculum, transfection conditions, amount and type of DNA used (including Filler DNA), and post-transfection culture conditions, we arrived at an uniquely optimized TGE process with higher titer and reduced costs and time, thus increasing the overall efficiency of early antibody material supply.