Flexible Microfluidic Process Technology for Biopharmaceutical Purification of Bacteriophages

Multidrug resistant (MDR) bacterial wound infections remain a persistent challenge for front-line military medical providers in prolonged care treatment. Phage therapeutics have demonstrated preclinical and clinical efficacy against ESKAPEE infections. Phage production however remains a challenge to remove bacterial remnants including LPS endotoxins and other pyrogens. To meet regulatory requirement for human applications, rapid and scalable phage purification technologies are urgently needed. In this project, we propose to develop a flexible and safe phage purification platform, comprising of rapid, efficient and scalable microfluidic technology to remove endotoxin (LPS) in phage preparations below safe levels for human use. In Phase I, we will develop a modular and automated purification platform, which uses label-free and size-based microfluidic separation technology to remove endotoxin. We will also develop real-time LPS measuring approach to allow automation and enhance product safety. In Phase II, we will focus on integration of system automation to decrease processing time and costs, as well as, test the performance for diverse array of Gram-negative phages. Phase III will focus on manufacturing alpha and beta prototypes. Our innovative Gram-negative phage purification platform will provide augmented capability in phage manufacturing for both military and civilian health applications.