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. Bacteriophage (phage) therapeutics have demonstrated preclinical and clinical efficacy against ESKAPEE infections. Phage production however remains a challenge to remove common pyrogen contaminants from phage products, including endotoxins (lipopolysaccharide (LPS) derived from Gram-negative bacteria), exotoxins, and non-endotoxic pyrogens. To meet safety requirement for human application, effective and scalable pyrogen removal technologies compatible for phage production are needed. Our overarching goal is to develop microfluidic filtration technologies for pyrogens removal from phage solutions that are rapid, efficient, and scalable. In Phase I, we prototyped a modular and automated endotoxin removal platform that employed size-based microfluidic separation of phage particles from endotoxin molecules. In addition, we developed a real-time endotoxin quantification device for platform automation and enhancement of phage product safety. In Phase II, we aim to integrate 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 of 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.