Protein composition and methods for analysing microbiota

Patent metadata

Field	Value
Publication number	US-11175294-B2
Application number	US-201716305461-A
Country	US
Kind code	B2
Filing date	Jun 1, 2017
Priority date	Jun 1, 2016
Publication date	Nov 16, 2021
Grant date	Nov 16, 2021

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 a plurality of microbiota samples from one or more human subjects, soil, or animals in culture;
spiking said culture of plurality of microbiota samples with an isotope-labelled standard, which comprises a microbiota from a given human, soil, or animal microbiota sample labelled with an isotope;
performing a metaproteomic analysis on the culture of plurality of microbiota samples and the isotope-labelled standard to identify changes in the microbiomes of said plurality of microbiota samples;
selecting said microbiomes exhibiting the identified changes; and
analyzing the selected microbiomes to characterize the changes.

2. The method as defined in claim 1 , wherein said provided plurality of microbiota samples are cultured in micro-well receptacles.

3. The method as defined in claim 2 , wherein said provided plurality of microbiota samples are cultured in micro-well plates.

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

5. The method as defined in claim 4 , wherein said microbial gene catalog of a given subject type is a microbial gene catalog of a human.

6. The method as defined in claim 4 , wherein said microbial gene catalog of a given subject type is a microbial gene catalog of an animal.

7. The method as defined in claim 1 , wherein said analyzing comprises performing a metaproteomic analysis combined with a metagenomic analysis.

8. The method as defined in 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 culture of plurality of microbiota samples.

9. The method as defined in 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.

10. The method as defined in 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.

11. The method as defined in claim 10 , wherein the given human microbiota sample is from an intestinal microbiota.

12. The method as defined in 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.

13. The method as defined in 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.

14. The method as defined in 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.

15. The method as defined in claim 1 , wherein the isotope comprises a stable isotope, a radioactive isotope, or both.

16. The method as defined in claim 15 , wherein the isotope comprises 13 C, 14 C, 15 N, 32 S, 35 S, 32 P, deuterium, or any combination thereof.

17. 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 a plurality of microbiota samples from one or more human subjects, soil, or animals;
spiking said plurality of microbiota samples with an isotope-labelled standard, which comprises a microbiota from a given human, soil, or animal microbiota sample labelled with an isotope;
performing a metaproteomic analysis on the plurality of microbiota samples and the isotope-labelled standard to identify changes in the microbiomes of said plurality of microbiota samples;
selecting said microbiomes exhibiting the identified changes; and
analyzing the selected microbiomes to characterize the changes.

Assignees

• Univ Ottawa

Inventors

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

Classifications

• A61B2503/42

  for laboratory research · CPC title

• G16B30/10

  Sequence alignment; Homology search · CPC title

• C12Q1/16 Primary

  using radioactive material · CPC title

• G01N33/6848 Primary

  Methods of protein analysis involving mass spectrometry · CPC title

• A61B5/4255

  Intestines, colon or appendix · CPC title