Bacteriophage replication on permissive host cells in fused silica capillary with nanostructured part as potential of electrophoretic methods for developing phage applications

Microscale electrokinetic‐based analysis of intact cells and viruses

“…The findings illustrated that 10 4 cells seemed to be the sweet spot, since using 10 3 host cells did not produce enough phages, and increasing the number of host cells to 10 5 hindered the success of the separation due to generation of aggregates and cell debris. Similar to their previous work [57], which required identifying appropriate BGE additives, selecting appropriate operating conditions proved to be essential to the success of the separation, which was carried in 20 min.…”

“…Since then, there has been significant progress in the development of effective and rapid electromigration‐based methods for the analysis of intact viruses. Of particular interest are techniques for clinical applications, such as the one reported Horká et al [57] in 2018 where they employed both CZE and IEF to study the bacteriophage strain K1/420 (from the Kayvirus genus), which is used for the therapy of MRSA. Phage therapy, which is the use of bacterial viruses (phages) to treat bacterial infections, is gaining increasing attention due to the growing number of antibiotic‐resistant bacteria.…”

“…An important challenge is the fact that the virus and the bacteria may have similar surface properties (hydrophobicity/hydrophilicity), since during infection the virus remains attached to host bacterial cells, which may provide enough time for their characteristics to become similar. Considering this challenge, Horká et al [57] focused first on separating K1/420 phages from the host bacteria. Figure 4A shows five distinct CZE electropherograms (labeled 1–5) illustrating the optimization of the separation of bacteriophage K1/420 purified by standard CsCl density gradient centrifugation process.…”

“… (A) CZE electropherograms of K1/420 purified by standard CsCl density gradient centrifugation process (electropherograms 1–5) and separation of a mixture of 5 × 10 9 PFU/mL of the phage spiked with 1 × 10 7 cell/mL of S. aureus bacteria (electropherogram #6). Adapted with permission from [57], copyright (2018) Elsevier. (B.i ) Schematic representation of separation, propagation, and characterization process designed by Horká et al.…”

“…(D.ii) CIEF electropherogram of the separation of 1 × 10 10 PFU/mL of the phage spiked with 1 × 10 7 cell/mL of S. aureus bacteria. Adapted with permission from [57], copyright (2018) Elsevier.…”