US 12,448,705 B2
Bacteriophage compositions and methods of selection of components against specific bacteria
James M Regeimbal, Washington's Crossing, PA (US); Biswajit Biswas, Germantown, MD (US); and Matthew S. Henry, Thurmont, MD (US)
Assigned to The Government of the United States of America, as Represented by the Secretary of the Navy, Arlington, VA (US)
Filed by United States of America as Represented by the Secretary of the Navy, Silver Spring, MD (US)
Filed on Aug. 16, 2022, as Appl. No. 17/820,035.
Application 16/434,320 is a division of application No. 15/628,368, filed on Jun. 20, 2017, granted, now 10,357,522, issued on Jul. 23, 2019.
Application 17/820,035 is a continuation of application No. 16/434,320, filed on Jun. 7, 2019, abandoned.
Claims priority of provisional application 62/510,649, filed on May 24, 2017.
Claims priority of provisional application 62/489,860, filed on Apr. 25, 2017.
Claims priority of provisional application 62/353,517, filed on Jun. 22, 2016.
Prior Publication US 2023/0079967 A1, Mar. 16, 2023
Int. Cl. C40B 40/02 (2006.01); C40B 30/04 (2006.01)
CPC C40B 40/02 (2013.01) [C40B 30/04 (2013.01); G01N 2500/10 (2013.01)] 23 Claims
OG exemplary drawing
 
1. A phage cocktail directed against a bacterial pathogen produced by a method comprising:
(a) constructing a bacterial diversity set comprising diverse strains of the same species as said bacterial pathogen, said constructing comprising collecting a plurality of bacterial isolates of the same species as said bacterial pathogen, analyzing said plurality of bacterial isolates to identify bacterial isolates which are clinically, genotypically and/or metabolically diverse strains of said species of bacterial pathogen, and down selecting said plurality of bacterial isolates to include said clinically, genotypically and/or metabolically diverse strains of said species of bacterial pathogen to create said bacterial diversity set;
(b) collecting mixed phages from diverse environmental sources;
(c) constructing a Tier 1 archival phage library, said constructing comprising hosting the phages collected in step (b) on one or more of the diverse strains of said species of bacterial pathogen comprising the bacterial diversity set created in step (a) in order to identify and purify lytic phages against strains of said bacterial species, and selecting said lytic phages for the Tier 1 archival phage library;
(d) constructing a Tier 2 working phage library, said constructing comprising characterizing the Tier 1 archival phage library constructed in step (c) to identify and exclude phages which demonstrate undesirable and/or toxic characteristics, further screening remaining phages in the Tier 1 working phage library against one or more of the diverse strains of said species of bacterial pathogen comprising the bacterial diversity set created in step (a) to characterize host range of each said remaining phages, and selecting phages free of undesirable and/or toxic characteristics and having desirable host ranges for the Tier 2 working phage library;
(e) screening the Tier 2 working phage library constructed in step (d) for individual phages and/or various phage combinations that may be therapeutically effective against the bacterial pathogen, said screening comprising performing phage efficacy assays, wherein said phage efficacy assays comprise growing cultures of said bacterial pathogen with individual phages, and/or various phage combinations from the Tier 2 working phage library, and analyzing bactericidal activity against said bacterial pathogen by said individual phages and/or said various phage combinations in said cultures, wherein a suitable delay in bacterial growth and/or a lack of appearance of phage-resistant bacterial growth in said cultures indicates said individual phages and/or said various phage combinations may be therapeutically effective against the bacterial pathogen; and
(f) compounding one or more of said individual phages, and/or said various phage combinations that may be therapeutically effective identified in step (e) to form said phage cocktail,
wherein the bacterial pathogen is susceptible to being killed by the phage cocktail.