Synergistic bacterial compositions and methods of production and use thereof

A technology of composition and bacteria, applied in the direction of bacteria, drug combination, medical raw materials derived from bacteria, etc., can solve problems such as difficulty in large-scale implementation and non-standardization

Active Publication Date: 2016-09-28
SERES HEALTH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, fecal transplantation is generally reserved for recurrent cases because of the potential for transmission of infectious or allergenic agents between hosts, the potential transfer of hundreds of unknown stra

Method used

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  • Synergistic bacterial compositions and methods of production and use thereof
  • Synergistic bacterial compositions and methods of production and use thereof
  • Synergistic bacterial compositions and methods of production and use thereof

Examples

Experimental program
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Effect test

example 1

[0244] Example 1. Construction of binary pairs in a high-throughput 96-well format / plate preparation

[0245] Bacterial pairs were used to identify binary pairs suitable for inhibiting C. difficile. To prepare plates for the high-yield screening assay of binary pairs, vials of the -80°C glycerol stock bacterial library were thawed and diluted to 1e8 CFU / mL. Each strain was then diluted 10X (to achieve a final concentration of 1e7 CFU / mL of each strain) into 200 [mu]L PBS + 15% glycerol in a well of a 96-well culture plate. Plates were then frozen at -80°C. When required, when tested with C. difficile in the CivSim assay, plates were removed from -80°C and thawed anaerobically at room temperature.

example 2

[0246] Example 2. Construction of triplets in a high-throughput 96-well format

[0247] Triple combinations of bacteria were used to identify triple combinations useful for inhibiting C. difficile. To prepare plates for high-yield screening triplet combinations, vials of the -80°C glycerol stock bacterial library were thawed and diluted to 1e8 CFU / mL. Each strain was then diluted 10X (to achieve a final concentration of 1e7 CFU / mL for each strain) into 200 μL of PBS + 15% glycerol in a well of a 96-well plate. Plates were then frozen at -80°C. When assay required, plates were removed from -80°C and thawed anaerobically at room temperature when tested with C. difficile in the CivSim assay.

example 3

[0248] Example 3. Construction of a CivSim analysis for screening bacterial compositions that inhibit the growth of Clostridium difficile

[0249] Compositions that inhibit the growth of C. difficile were identified using a competition assay (CivSim assay). Briefly, overnight cultures of C. difficile were grown in SweetB-FosIn under anaerobic conditions to allow C. difficile to grow. In some cases, other suitable media can be used. SweetB-FosIn is a version of BHI (Remel R452472) supplemented with the following components: Components per liter: 37g BHI powder (Remel R452472), supplemented with 5g yeast extract UF (Difco210929), 1g cysteine-HCl ( Spectrum C1473), 1g cellobiose (Sigma C7252), 1g maltose (Spectrum MA155), 1.5ml hematin solution, 1g soluble starch (Sigma-Aldrich S9765), 1g fructooligosaccharide / inulin (Jarrow Formulas 103025) and 50 mL 1M MOPS / KOH pH 7. To prepare the hematin solution, hematin (Sigma 51280) was dissolved in 0.1 M NaOH to make a 10 mg / mL stock. ...

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Abstract

Provided are therapeutic compositions containing microbial populations for prevention, treatment and reduction of symptoms associated with a dysbiosis of a mammalian subject such as a human.

Description

[0001] Related applications [0002] This application is related to commonly owned International Application No. PCT / US 13 / 2013, entitled "SYNERGISTIC BACTERIAL COMPOSITIONS AND METHODS OF PRODUCTION AND USETHEREOF", filed on November 25, 2013 71758, which both claim U.S. Provisional Patent Application No. 61 / 729,518, filed Nov. 23, 2012, U.S. Provisional Patent Application No. 61 / 729,519, filed Nov. 23, 2012, Nov. 23, 2012 Pending U.S. Provisional Patent Application No. 61 / 729,520, U.S. Provisional Patent Application No. 61 / 729,521, filed November 23, 2012, U.S. Provisional Patent Application No. 61 / 729,522, filed November 23, 2012 , U.S. Provisional Patent Application No. 61 / 729,524 filed on November 23, 2012, U.S. Provisional Patent Application No. 61 / 729,515 filed on November 23, 2012, and U.S. Provisional Patent Application No. 61 / 729,515 filed on November 23, 2012 Application No. 61 / 729,517, U.S. Provisional Patent Application No. 61 / 729,525, filed November 23, 2012, U.S...

Claims

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Application Information

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IPC IPC(8): A61K35/742A61K35/744A61K35/745A61K35/00
CPCA61K35/74A61K2035/11A61K2035/115C12N1/20A23L33/135A61P1/04A61P1/14Y02A50/30A61K45/06A61K35/742A61P1/00
Inventor M·R·赫恩K·D·利特科夫斯基A·M·德奥诺弗里奥T·K·奥苏米M-J·L·麦肯齐G·冯马尔扎恩D·N·库克D·A·贝利N·B·阿菲延J·G·奥宁斯
Owner SERES HEALTH
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