Fast Monitoring of Bacteriophage Production During Fermentation Using the Agilent Bio-Monolith HPLC Column

Bacteriophages, viruses that infect bacteria, were discovered at the beginning of the twentieth century. Today, phages are being used as antibacterial agents, in phage display screening, in the development of phage delivery vaccines, as targeted gene therapy delivery systems, and for specific bacteria typing. In order to use phages in these applications, they are commonly purified by liquid chromatography and must be free of all impurities. A purification and concentration process was recently developed using an ion exchange monolithic column [1]. After propagation and lysis of their host, phages are released into the growth medium. Along with the phages, other host components are released into the growth media, one of them being the host genomic DNA (gDNA), which poses a problem because it soon starts to degrade and form smaller fragments. When purifying phages with ion-exchange chromatography, it is crucial that the medium containing the phages does not contain any fragmented DNA. Free DNA fragments can interfere in the in vitro phage applications, phage display, and bacteria typing, and can be toxic in the therapeutic phage applications, vaccine production, and gene delivery. The Agilent Bio-Monolith DEAE column (weak anion exchange) can be used to monitor the fermentation process and to evaluate the amount of degraded gDNA throughout the purification process. Authors Franci Smrekar, Aleš Podgornik, Mateja Ciringer, and Lidija Urbas BIA Separations Teslova 30 Sl-1000 Ljubljana Slovenia Peter Raspor Department of Food Science and Technology, Biotechnical Faculty University of Ljubljana Jamnikarjeva 101 SI-1000 Ljubljana Slovenia Taegen Clary Agilent Technologies, Inc. 5301 Stevens Creek Blvd. Santa Clara, CA 95052 USA Application Note