Phage survival: The biodegradability of M13 phage display library in vitro

Abstract

Administration of bacteriophages is used for phage therapy modulation of gut microbiome or for in vivo phage display. The aim of the study was to analyze the survival of M13 phage in different body fluids and tissues in vitro. The survival of M13 phage was measured in vitro in human blood, saliva, urine, artificial gastric juice (AGJ), and mouse homogenates of stomach, jejunum, and colon after defined time points (5, 15, or 45 Min). The plates were inspected after overnight incubation and the plaques were counted. No phage was recovered after 5 Min of incubation with AGJ. In urine, the phage survival was decreased by 44% after 5 Min of incubation (P = 0.004). In saliva, the recovered titer was decreased by 33% and 88% (P < 0.05) after 15 and 45 Min, respectively. Phage coincubation with jejunum homogenate led to significant decrease of phage titer by 72% (P < 0.01) after 15 Min and by 99% (P < 0.001) after 45 Min. Decreased survival of M13 phage depending on time of incubation was proved under several in vitro conditions, with low pH in the AGJ having the most detrimental effect on phage survival. Phage pharmacokinetics described in vitro might have applications for the use of bacteriophages in vivo.

A bacteriophage cocktail targetingEscherichia colireducesE. coliin simulated gut conditions, while preserving a non-targeted representative commensal normal microbiota

“…This study's results diverge from the study by Ma et al., who reported larger (1.29 and 1.67 log units, respectively) loss of Salmonella phage Felix O1) titers in 1% and 2% bile solution. 27 The negative impact of bile salts on phage viability has been reported by some other authors as well 28-30 ; however, the topic is still not fully understood as other investigators – and this study – suggest that phages can be fairly stable and remain infectious when exposed to bile salts ranging from 4–10 mM). 31 It is possible that the phage viability will be more profoundly reduced in vivo due to many additional factors (e.g., intestinal peristalsis, more complex microbiota, various diets, etc.)…”

The use of a salmonella bacteriophage in bearded dragons: application, passage time and reisolation

“…As phages could be reisolated up to 24 days after the last application, they must have successfully reproduced, therefore have lysed salmonella to replicate themselves, because there is no evidence in literature that a Felix O1 phage is capable of surviving in a gastrointestinal tract for longer periods, without reproducing itself. Tóthová et al [34] designed a mouse homogenate of jejunum and colon and incubated these with a M13 phage without presenting the natural host of this phage. They demonstrated a significant reduction of phage activity after 45 minutes.…”

SMV1, an extremely stable thermophilic virus platform for nanoparticle trafficking in the mammalian GI tract

“…In order to evaluate SMV1 against a commonly used viral nanoplatform, we compared it to the E. coli bacteriophage, M13KE, which is the most commonly used phage for in vivo phage display experiments (Tothova et al . ) and as an addressable nanoprotein cage architecture (Yi et al . ).…”

“…Sulfolobus monocaudavirus 1 extremophilic nature suggests it can survive the harsh conditions present in the GI tract of humans and animals. In order to evaluate SMV1 against a commonly used viral nanoplatform, we compared it to the E. coli bacteriophage, M13KE, which is the most commonly used phage for in vivo phage display experiments (Tothova et al 2012) and as an addressable nanoprotein cage architecture (Yi et al 2012). Sulfolobus monocaudavirus 1 showed no decline in infectivity when incubated in 50% DMSO.…”

“…Viral stability in simulated GI conditions is valuable for prediction of in vivo behaviour of drug formulations (Klein ) and potential viral nanoplatform‐based vectors, like phages (Tothova, Babickova, and Celec ). Two types of media were used to evaluate the stability of SMV1 particles: SIF (simulation of small intestinal conditions in the fasted state), and SGF (simulation of stomach conditions in the fasted state).…”