SBIR-STTR Award

The manufacturing of recombinant protein-based biopharmaceuticals is a complex, labor and capital intensive endeavor. It currently is accepted that the use of mammalian cells is a requirement for the production of most human proteins, which typically contain extensive post-translational modifications that are not performed by prokaryotes and single-celled eukaryotes. Although mammalian cells such as Chinese hamster ovary cells and baby hamster kidney cells can faithfully biosynthesize most human proteins, the efficiency is dramatically lower than is achieved by bacterial or yeast cells. Of the recombinant proteins currently marketed, coagulation factor VIII is manufactured with the lowest efficiency and is by far the most expensive on a per unit mass basis. Other expensive and difficult to manufacture biopharmaceuticals include the other coagulation factors (e.g. factors IX and VIIa), interferon, and hormone-based cytokines. The goal of the current application is to develop a technology that can improve the production volume and pharmacy price of these difficult to manufacture recombinant biologics. Expression Therapeutics has developed a new baby hamster kidney-derived (BHK) cell line, designated BHK-MS, which can be cultured and expanded indefinitely in serum and blood product-free medium under suspension bioreactor conditions. In the current studies, we are 1) developing and validating the suspension adapted BHK-MS cell line for superior coagulation factor VIII and factor IX production, and 2) performing a pilot-scale manufacturing run and analyzing the biochemical and pharmaceutical characteristics of BHK-MS biosynthesized recombinant factor VIII and factor IX, which will serve as a model, difficult to manufacture biopharmaceuticals.

Public Health Relevance:
The long-term commercial objective of the current project is to commercialize a suspension cell-based biomanufacturing platform designed for improved volumetric productivity of complex, hard to manufacture biologics. Low-yield manufacturing processes currently in place often result in limited product availability and high-product costs. Expression Therapeutics has developed a patent-pending cell line designed for efficient manufacturing of recombinant biologics. The studies proposed herein are designed to provide proof-of-concept of the efficiency and scalability, as well as, rigorous preclinical validation and characterization of this novel manufacturing platform.

Public Health Relevance Statement:
The long-term commercial objective of the current project is to commercialize a suspension cell-based biomanufacturing platform designed for improved volumetric productivity of complex, hard to manufacture biologics. Low-yield manufacturing processes currently in place often result in limited product availability and high-product costs. Expression Therapeutics has developed a patent-pending cell line designed for efficient manufacturing of recombinant biologics. The studies proposed herein are designed to provide proof-of-concept of the efficiency and scalability, as well as, rigorous preclinical validation and characterization of this novel manufacturing platform.

NIH Spending Category:
Bioengineering; Biotechnology; Hematology; Kidney Disease

Project Terms:
A Mouse; Address; Adolescence; Amino Acids; base; Biochemical; Biological Products; Biological Response Modifier Therapy; Biomanufacturing; Biomedical Engineering; Bioreactors; Blood Coagulation Factor; blood product; Blood Proteins; Capital; Cell Aggregation; Cell Culture Techniques; Cell Density; Cell Line; Cells; Characteristics; Chinese Hamster Ovary Cell; Clinical; Column Chromatography; Complex; Conditioned Culture Media; Consumption; cost; cytokine; design; Development; Disease; Drug Formulations; Electroporation; Eukaryota; Evaluation; Factor IX; Factor VIII; Family suidae; flasks; flexibility; Generations; Genetic; genetically modified cells; Goals; Gold; Growth; Hamsters; Harvest; Hemophilia A; Hormones; Human; improved; in vivo; Interferons; Ion Exchange; Kidney; kidney cell; Kilogram; Legal patent; Lentivirus Vector; Liposomes; M cell; Mammalian Cell; manufacturing process; Marketing; Measures; Mediating; Methods; Modeling; Modification; Monoclonal Antibodies; Mortality Vital Statistics; novel; Nutrient; Persons; Pharmacologic Substance; Pharmacy facility; Phase; Polymers; Post-Translational Protein Processing; pre-clinical; Price; Procedures; Production; Productivity; Prokaryotic Cells; Protein Biosynthesis; Proteins; recombinant antihemophilic factor VIII; Recombinant Proteins; Recombinants; Relative (related person); research and development; Roller Bottle; Running; Serum; Serum Proteins; Serum-Free Culture Media; Small Business Innovation Research Grant; Suspension Culture; Suspension substance; Suspensions; System; Technology; Testing; Therapeutic; Thrombin; Time; Validation; Yeasts

The long-term objective of current project is to commercialize a novel coagulation factor product manufactured using a novel production cell line, designated ET3i/BHK-M with a primary indication of treatment of anti-drug antibodies that develop following the treatment of congenital hemophilia A with factor VIII (fVIII) replacement products. Expression Therapeutics has developed multiple technologies that, when integrated, provide a highly efficient biomanufacturing platform designed for improved volumetric productivity. In the phase I SBIR project periods, Expression Therapeutics generated and validated a research cell bank demonstrating the highest levels of recombinant factor VIII reported to date, completed upstream and downstream process development, and successfully transferred the technology to a contract manufacturing organization for production of phase I clinical test material. The current application seeks to complete all manufacturing, nonclinical, clinical planning, and regulatory activities required to conduct a phase I first-in-man trial of ET3i/BHK-M, with the primary endpoint of safety and tolerability.

Public Health Relevance Statement:
PROJECT NARRATIVE Despite many advances toward the control and prevention of bleeding in hemophilia A through coagulation factor VIII infusion therapy, 25 – 35% of severe patients develop anti-drug antibodies termed ‘inhibitors’ that block treatment efficacy. In the US, Canada and Western Europe, the majority of inhibitor patients are treated off-label with higher, more frequent administration of expensive coagulation factor VIII products in protocols collectively referred to as ‘Immune Tolerance Induction’ therapy. The goal of the current project is to commercialize the first product using our novel commercial cell production system, which will be a coagulation factor VIII product with a primary label indication for the treatment of fVIII inhibitors through Immune Tolerance Induction therapy.

Project Terms:
Antibodies; barrier to care; Biological Response Modifier Therapy; Biological Sciences; Biomanufacturing; Biomedical Engineering; Bioreactors; Blood Coagulation Factor; Businesses; Canada; Capital; Caring; cell bank; Cell Line; Cells; Chemistry; Clinical; clinical development; clinical material; commercialization; Complication; Contracts; cost; Country; Data Analyses; design; Development; Development Plans; Documentation; Dose; Economics; F8 gene; Factor VIII; Family suidae; Feedback; first-in-human; Goals; Half-Life; Hamsters; Hemophilia A; Hemorrhage; Human; Immune Tolerance; improved; Infusion procedures; inhibitor/antagonist; innovation; Insurance; International; Investments; Iowa; Kidney; Label; Laboratories; Legal patent; M cell; Materials Testing; meetings; Methods; Multicenter Trials; National Heart, Lung, and Blood Institute; Neoadjuvant Therapy; novel; open label; Patients; Persons; Pharmaceutical Preparations; Pharmacology and Toxicology; Phase; Policies; Population; preclinical development; Prevention; primary endpoint; Process; Production; Productivity; programs; Proteins; Protocols documentation; R-factor; recombinant antihemophilic factor VIII; Recombinants; Records; Recovery; Reporting; Research; research clinical testing; Resources; Running; Safety; safety study; Serum; Services; Small Business Innovation Research Grant; stability testing; Suspensions; System; Technology; Technology Transfer; Teenagers; Testing; Therapeutic; therapy design; Treatment Efficacy; Treatment Factor; Western Europe; Work