Sabin and IBT have successfully produced MARV rGP needed to support all clinical and non-clinical testing and the material is now being used in a fit-for-use assay for a Phase Ib human participants plasmapheresis clinical trial. Qualification and validation can begin for upcoming non-human primate and human sample testing from Sabin's Marburg Vaccine (Investigational Monovalent Chimpanzee Adenoviral Vectored Filovirus) studies to measure IgG levels and thereby identify Marburg GP IgG as a correlate of protection in order to demonstrate potential efficacy in humans. Funding under BARDA contract HHSN 75A50119C00055 is acknowledged.

Half-life extension of APIs has proven to be an effective way to treat patients using lower doses or less frequent dosing. Technologies to increase product half-life have evolved, introducing new challenges for traditional purification strategies. In this presentation, we share the effects of charge-shielding attributed to genetic fusion of a proline and alanine repeat polypeptide to a therapeutic enzyme, and the development of a non-traditional approach to a purification process.

The success of Protein A affinity capture for mAbs has driven a search for similar methods for other proteins. The challenge has been designing affinity resins for arbitrary products with no common features. In this talk, I will compare and contrast three emerging solutions: custom affinity resins designed for single products, cleavable tags that rely on proteases, and self-removing tags that rely on inteins.

Due to the costly and time-consuming production of biologicals, selective affinity purification methods are desirable. For efficient and selective purification, a number of criteria are to be met. One important feature is the durability and possibility to reuse the purification matrix. Another is the possibility to avoid harsh conditions during the purification scheme. Different strategies to address these issues when developing affinity purification systems will be presented.

In search of the next-generation chromatographic technique for antibody purification, our team has developed an affinity chromatography method using the unconventional nucleotide binding site (NBS). By generating resins that display ligands that bind to the NBS that is conserved on all immunoglobulins, we achieve purity over 99% with >99% column efficiency, supporting that NBS method provides a stable, reusable, and inexpensive alternative for purification of humanized and chimeric antibodies.

Immobilized metal affinity chromatography (IMAC) is the workhorse of a majority of recombinant protein purification. However, one-third to one-half of all proteins are considered metalloproteins. This begs consideration that IMAC purification may cause problems. Therefore, we designed a new vector that replaces the His-tag with a streptavidin-based affinity tag, TwinStrep. We cloned into this vector, KRAS4b G-domain, PIK3ca RAS binding domain (RBD), and RAF1 RBD plus its cystine-rich domain (CRD). KRAS4b and PIK3ca were indistinguishable, but RAF1 showed significant improvement in folding suggesting that metal-free purification produced a protein conformation more similar to its native fold.

A truly continuous biomanufacturing process will be presented. This is achieved by integration of a perfusion bioreactor with continuous precipitation and continuous filtration, replacing the state-of-the-art capture step for mAbs, protein A affinity chromatography. Surge tanks have been omitted in lieu of tubular reactors. Hereby, the residence time and the start-up/shut-down are very short, the residence time distribution very narrow. Yield, purity, and critical quality attributes are shown.

Protein production is more complex than just the act of expressing the protein. This presentation will review the end-to-end protein production workflow process and reflect on possibilities of how to increase the efficiency and productivity of a recombinant protein expression facility.

X-ray free electron lasers probe protein structures with ultrashort x-ray pulses thereby enabling the determination of molecular movies of molecules “at work," but large quantities of proteins in the range of hundreds of milligrams are required. We will present strategies and procedures for large-scale cell culture and protein isolation for XFEL studies that include preparation of large photosynthetic membrane protein complexes, as well as preparation of proteins from SARS-CoV-2. 

High-throughput protein production relies on affinity tags to enable purification of new targets, where tags are often left in place during initial target characterization. The production of tagless targets is now possible via a self-removing tag that functions in simple buffer systems. We will describe the use of this system to purify a variety of targets to extremely high purity with a standard protocol on a universal affinity resin.

As biologic therapeutics continue to increase in complexity, innovative approaches to candidate screening, production, characterization, and development are more important than ever. Our advanced protein production workflows incorporate novel processes, intelligent high-throughput automation, and high-quality informatics to enable robust molecule screening, selection, and scale-up. These enhancements enable advances in the speed, quality, and productivity of our biologics development pipeline.

Drug Discovery landscape is ever evolving and constantly demands revolutionary advancements in protein expression and production core laboratories. This presentation will focus on the evolution of our end-to-end automated high-throughput protein expression, purification workflows, and building creative solutions to support ever-increasing demands in today's world. It will emphasize the importance of collaboration and integration between different sub-groups including molecular biology, automation, and bioinformatics to provide more efficient processes. These multi-facet approaches have significantly alleviated some of the bottlenecks in protein production and accelerates the provision of key protein reagents to ambitious projects, particularly for challenging and undruggable targets.

Join a Think Tank discussion group to share and experience and hear what others have learned.
1) Workflow vs technology development?
2) What might address the future and what is needed?
3) Issues and challenges with end-to-end protein production?
4) Tearing down silos – how do you foster cross-functional collaborations to innovate and improve
5) Takeaways from PepTalk: The Protein Science and Production Week?