Enabling Next-Generation Biomanufacturing Using Cell-Free Technology

Weston Kightlinger, director, cell-free protein synthesis, National Resilience.

Kightlinger shared his company’s work to enable next-generation biomanufacturing, focusing on biologics and cell-free protein synthesis. Resilience was formed to address the need for robust manufacturing supply chains that remain stable amid times of disruption and enable design of therapeutics in pace with scientific advancements. Resilience is a technology-focused manufacturing company dedicated to democratizing access to complex medicines. It is building a network of high-tech manufacturing facilities: 11 sites, 1,700 employees, and a million feet of manufacturing. The focus is to “boot up” standardized and multimodal networks so that there is no single point-of-failure across the supply chain, with end-to-end customer transparency and reduced wait times, shorten times to clinic, and faster delivery to patients.

Resilience has offerings across a number of modalities. The biologics unit is dedicated to meeting industry needs for speed to clinic through innovative technology and an end-to-end platform from gene to IND. The company uses established technology and advanced technology such as continuous manufacturing and cell-free processes. It continues to build capacity toward supporting commercially approved facilities. The Resilience biologics network has expertise in a broad range of molecules, from monoclonal antibodies (MAbs) to enzyme, bispecifics, and other complex proteins in processes using fed-batch and perfusion cultures and a number of cell lines. It hopes to “widen the aperture” of biological systems to address the challenge of increasingly complex therapeutics. Kightlinger explained benefits of cell-free synthesis that enable production of a large number of protein therapeutics.

Resilience is dedicated to building the expertise needed to customize cell-free reactions (“modules”) to produce different types of therapeutics. Kightlinger presented key ways in which Resilience is accelerating cell-free biomanufacturing. He also outlined synergy with the company’s technology ecosystem, integration into the manufacturing infrastructure, and internal and external company engagement across modalities in the drug development process to make the most of the cell-free process.

Difficult-to-manufacture proteins are being unlocked through cell-free production, through use of a cell-free chassis strain for engineering, and by enabling cell-free glycoprotein synthesis. Kightlinger explained his company’s success in customizing its cell-free process for a number of protein classes: antibodies, multispecifics, fusion proteins, various types of enzymes, subunit vaccines, posttranslationally modified proteins, and cytotoxic and other complex proteins.

A case study illustrated successful expression of a challenging protein motif: a complex cystine knot for a compelling acute indication. A specific formulation of Resilience cell-free reaction components was able to unlock it for scalable manufacturing, with a titer increase of over 100-fold with strong expression of all nine of the constructs within the therapeutic class.

Kightlinger shared general strategies, noting that because one strain does not fit all, Resilience continues to innovate and design its chassis strains for ideal production. It is generating a library of strains to apply to unique types of therapeutics with unique production needs. He highlighted the company’s approach to enabling scalable cell-free glycoprotein synthesis through expression of a lipid-linked oligosaccharide plasmid for eventual integration into the genome. Resilience is building the glycans and a critical enzyme catalyst for transfer onto a protein within the cell-free reaction, accomplishing coupled transcription–translation and glycosylation all in one step, which is far more scalable than adding extra components to the reaction.

Synergies with the company’s biomanufacturing technology ecosystem provide continuous GMP manufacturing as a core competency. Other integrated functions include CMC, QA, and regulatory affairs. A major focus of the company from its inception has been supply-chain management.

Plans to build out a number of applications and synergies across modalities will provide biomanufacturing as a service, responding to defense and pandemic needs. Cell-free synthesis does not require development of unique CHO cell lines, and it scales linearly because it’s a biochemical reaction, thus bypassing concerns about cell rupture and cell viability that often provide difficulties during scale-up.

Fill out the form below to view the full BPI Theater presentation.