(443c) Toward on-Demand Personalized Treatment of Acute Lymphoblastic Leukemia through Low-Cost at-Home Cell-Free Protein Synthesis Biosensors

Unfortunately, thousands of children in the US are diagnosed each year with Acute Lymphocytic Leukemia (ALL), the most common form of childhood cancer. The FDA approval and widespread adoption of treatment with bacterial asparaginase (ASNase) greatly improved survival rates in the 1960-1980s to ~80% where they remain today. However, during treatment there is a high incidence of immune system neutralization of ASNase and dosage is currently not personalized. In recent studies, survival rates of ALL increase 10% to ~90% when ASNase levels are consistently monitored for personalized treatment and detection of treatment neutralization events. Unfortunately, this is not the standard of care and requires many blood draws and specialized equipment for testing. To improve survival rates here we present the development of a low-cost at-home sensor that detects ASN levels in a drop of human blood (similar quantity currently required for the common glucose meter). The sensor is powered by an E. coli-based cell-free protein synthesis system. This presentation covers key advancements in the technology which include: 1) the efficient lyophilization of E. coli lysate with energy and DNA template molecules as a “single-pot” for a “just-add sample” sensor that is shelf-stable for months, 2) the development of effective low-cost techniques to separate red blood cells for visual analysis, 3) the engineering of low-cost production methods for RNAse inhibitor to maintain biosensor affordability (reduces cost by 90%), and 4) the utility of diverse reporter systems for rapid and accurate visual outputs with a cell-phone camera^1-5. Overall this work demonstrates the potential of an at-home glucose-meter-like sensor to enable personalized treatment and improve survival rates for ALL patients.

References:

1) Tucker RW, Free TJ, Simonson KM, Smith SA, Lindgren CM, Pitt WG, Bundy BC. 2023. Engineering At-Home Dilution and Filtration Methods to Enable Paper-Based Colorimetric Biosensing in Human Blood with Cell-Free Protein Synthesis. Biosensors. 13(1): 104.

2) Smith SA, Lindgren CM, Ebbert LE, Free TJ, Nelson JA, Simonson KM, Hunt JP, Bundy BC. 2023. “Just Add Small Molecules” Cell-Free Protein Synthesis: Combining DNA Template and Cell Extract Preparation into a Single Fermentation. Biotechnology Progress. https://doi.org/10.1002/btpr.3332

3) Soltani M, Bundy BC. 2022. Streamlining Cell-free Protein Synthesis Biosensors for Use in Human Fluids: In situ RNase Inhibitor Production during ExtractPpreparation. Biochemical Engineering Journal. 177:108158

4) Soltani M, Hunt JP, Bundy BC. 2021. Rapid RNase Inhibitor Production to Enable Low-cost on-Demand Cell-Free Protein Synthesis Biosensor use in Human Body Fluids. Biotechnology and Bioengineering. 118(10):3973-3983.

5) Hunt JP, Wilding KM, Barnett RJ, Robinson H, Soltani M, Cho JE, Bundy BC. 2020. Engineering Cell-Free Protein Synthesis for High-Yield Production and Human Serum Activity Assessment of Asparaginase: Toward On-Demand Treatment of Acute Lymphoblastic Leukemia. Biotechnology Journal. 5(4):e1900294.