Protein composition and methods for analysing microbiota

Patent metadata

Field	Value
Publication number	US-11933790-B2
Application number	US-202117159731-A
Country	US
Kind code	B2
Filing date	Jan 27, 2021
Priority date	Jun 1, 2016
Publication date	Mar 19, 2024
Grant date	Mar 19, 2024

Abstract

A method of isotope-labelling a microbiota sample. It involves providing a first microbiota sample that was obtained from a given source; exposing the first microbiota sample to an isotope enriched medium; and culturing the exposed first microbiota sample in the isotope enriched medium to obtain an isotope-labelled microbiota sample, wherein the isotope labelled metaproteome of the isotope-labelled microbiota sample is taxon specific for taxa present in the first microbiota sample when initially obtained from the given source.

First claim

What is claimed is:

1. A method of high throughput screening of multiple microbiota samples from a human subject, soil, or animal for metaproteomic analysis of said samples comprising:
providing proteome extracts from a plurality of microbiota samples from one or more human subjects, soil, or animals in culture;
spiking said proteome extracts with an isotope-labelled standard, which comprises a proteome of microbiota from a given human, soil, or animal microbiota sample labelled with an isotope by culturing said microbiota from a given human, soil, or animal microbiota in an isotope enriched medium;
performing a pre-screening using a metaproteomic analysis to identify changes in microbiomes of said plurality of microbiota samples;
selecting said microbiomes exhibiting changes; and
analyzing the selected microbiomes to characterize the changes.

2. The method of claim 1 , wherein said analyzing comprises using a microbial gene catalog of a given subject type and an iterative database search strategy.

3. The method of claim 2 , wherein said microbial gene catalog of a given subject type is a microbial gene catalog of an animal or human.

4. The method of claim 1 , wherein said analyzing comprises performing a metaproteomic analysis combined with a metagenomic analysis.

5. The method of claim 1 , wherein said spiking comprises adding sufficient isotope-labelled standard to reach a 1:1 protein mass ratio with the protein contained in said proteome extracts from the plurality of microbiota samples.

6. The method of claim 1 , wherein the plurality of microbiota samples are from one or more human subjects or animals, and wherein the method further comprises assessing the results of said analysis of the selected microbiomes to perform at least one of:
disease diagnosis in a target human subject or an animal;
assessing treatment response in a target human subject or an animal;
assessing remission in a human subject or an animal receiving treatment;
screening for xenobiotic effects on a microbiome of a target human subject or an animal;
screening for effects of a compound on a microbiome of a target human subject or an animal, wherein said compound is one of a food, a drug, a chemical, a therapeutic agent, a toxin, a poison, a beverage, a food additive, a cosmetic, a cosmetic ingredient, packaging material, a pesticide, a herbicide, a consumer product; and
screening a microbiome to identify the responsiveness of a human subject or an animal to a therapy or treatment.

7. The method of claim 1 , wherein said isotope-labelled standard comprises labelled proteins representative of a metaproteome from the given human, soil, or animal microbiota sample and having at least 90% average heavy isotopic enrichment rate.

8. The method of claim 7 , wherein the given human microbiota sample is from an intestinal microbiota.

9. The method of claim 1 , wherein said isotope-labelled standard is taxon-specific for at least about 50% of the microbe populations present in the plurality of microbiota samples from the one or more human subjects, soil, or animals.

10. The method of claim 1 , wherein said isotope-labelled standard is taxon-specific for at least about 85% of the microbe populations present in the plurality of microbiota samples from the one or more human subjects, soil, or animals.

11. The method of claim 1 , wherein the plurality of microbiota samples from the one or more human subjects are from an intestinal microbiota, a vaginal microbiota, an oral microbiota, a cutis microbiota, a bladder microbiota, a kidney microbiota, a lung microbiota, an eye microbiota, a breast microbiota, a penile microbiota, or any combination thereof.

12. The method of claim 1 , wherein the isotope comprises a stable isotope, a radioactive isotope, or both.

13. The method of claim 12 , wherein the isotope comprises 13 C, 14 C, 15 N, 32 S, 35 S, 32 P, deuterium, or any combination thereof.

Assignees

• Univ Ottawa

Inventors

• Figeys Joseph Michel Daniel
• Stintzi Alain Christophe
• Mack David R
• Zhang Xu
• Ning Zhibin

Classifications

• G01N33/6848 Primary

  Methods of protein analysis involving mass spectrometry · CPC title

• A61B5/4255

  Intestines, colon or appendix · CPC title

• C12Q1/16 Primary

  using radioactive material · CPC title

• G01N33/5023

  on expression patterns · CPC title

• G01N33/5091

  for testing the pathological state of an organism · CPC title