Point-of-Care Low-Cost Cancer Therapeutic Production and Monitoring Efficacy in Human Serum with Cell-Free Protein Synthesis: Toward on‐Demand Treatment of ALL

Acute lymphocytic leukemia (ALL) is the most common form childhood cancer. Fortunately, administration of bacterial asparaginase (ASNase) has greatly improved survival rates, however these therapeutics have high incidence of immunological neutralization and serum activity must be monitored for most effective treatment regimens. Here, improvement in cell‐free protein synthesis (CFPS) of FDA approved l‐asparaginase (crisantaspase) is reported by employing an aspartate‐fed‐batch reactor format. A CFPS‐based ASNase activity assay as a tool for therapeutic regimentation and production quality control is also presented. This work suggests that shelf‐stable and low‐cost Escherichia coli‐based CFPS reactions may be employed on‐demand to 1) synthesize biologics on‐site for patient administration, 2) verify biologic activity for dosage calculations, and 3) monitor therapeutic activity in human serum during the treatment regimen. This work demonstrates the potential of CFPS as both a production platform and biosensor for treatment and treatment monitoring in the clinical setting.

Reference: Hunt JP, Wilding KM, Barnett RJ, Robinson H, Soltani M, Cho JE, Bundy BC. Engineering Cell-Free Protein Synthesis for High-Yield Production and Human Serum Activity Assessment of Asparaginase: Toward On-Demand Treatment of Acute Lymphoblastic Leukemia. Biotechnol J. 2020. 15(4):e1900294.