Screening of Hydrophilic Polymers Reveals Broad Activity in Protecting Phages During Cryopreservation

Bacteriophages have many biotechnological and therapeutic applications, but as with other biologics, cryopreservation is essential for storage and distribution. Macromolecular cryoprotectants are emerging for a range of biologics, but the chemical space for polymermediated phage cryopreservation has not been explored. Here we screen the cryoprotective effect of a panel of polymers against 5 distinct phages, showing that nearly all the tested polymers provide benefit. Exceptions were poly(methacrylic acid) and poly(acrylic acid), which can inhibit phage-infection with bacteria, making post-thaw recovery challenging to assess. A particular benefit of a polymeric cryopreservation formulation is that the polymers do not function as carbon sources for the phage hosts (bacteria) and hence do not interfere with post-thaw measurements. This work shows that phages are amenable to protection with hydrophilic polymers and opens up new opportunities for advanced formulations for future phage therapies and to take advantage of additional functionality brought by the polymers.