Analyte enrichment methods and compositions

Patent metadata

Field	Value
Publication number	US-12559785-B2
Application number	US-202016810347-A
Country	US
Kind code	B2
Filing date	Mar 5, 2020
Priority date	May 13, 2013
Publication date	Feb 24, 2026
Grant date	Feb 24, 2026

Abstract

Methods and compositions for enriching a population of particles containing an analyte are disclosed. In one embodiment, enrichment beads are used that are larger in size than the beads used for amplification. A separation device is employed that can retain larger beads with bound amplified beads. The technique finds many uses, including enriching for beads with clonally amplified template, which can be used in a variety of assays, including nucleic acid sequencing.

First claim

The invention claimed is:

1 . A population of emulsion bead—enrichment bead complexes captured upon a single layer nylon mesh, said emulsion beads comprising amplified template, wherein said amplified template is amplified from one nucleic acid template, wherein said emulsion beads are non-covalently bound to said enrichment beads and wherein at least 50% of said emulsion beads comprise homogeneous amplified template.

2 . The complexes of claim 1 , wherein said emulsion beads are 1 μm in diameter.

3 . The complexes of claim 1 , wherein said population of emulsion beads comprises between 1 bead and 20 million beads.

4 . The complexes of claim 1 , wherein at least 80% of said population of emulsion beads comprises amplified homogeneous template.

5 . The complexes of claim 4 , wherein said population of emulsion beads is contaminated with less than 1% of beads without amplified template.

6 . The complexes of claim 1 , wherein said enrichment beads are larger than said emulsion beads.

7 . The complexes of claim 6 , wherein said emulsion beads are 1 μm in diameter and said enrichment beads are approximately 15 μm in diameter.

8 . The complexes of claim 1 , wherein said amplified template containing at least one biotinylated nucleotide comprises a first part of an interaction pair, and wherein said enrichment beads comprise the second part of said interaction pair.

9 . The complexes of claim 1 , wherein said enrichment beads comprise streptavidin and said amplified template of said emulsion beads comprises at least one biotinylated nucleotide, such that said complexes comprise a biotin streptavidin interaction.

10 . The complexes of claim 6 , wherein said enrichment beads are at least five times larger than emulsion beads.

Assignees

• Qiagen Sciences Llc
• Qiagen Sciences Inc

Inventors

• Olejnik Jerzy
• Gordon Steven
• Werner Martina

Classifications

• C12Q2563/155

  Particles of a defined size, e.g. nanoparticles · CPC title

• C12Q2563/149

  Particles, e.g. beads · CPC title

• C12Q1/6844

  Nucleic acid amplification reactions · CPC title

• C12Q1/6806 Primary

  Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay (C12Q1/6804 takes precedence) · CPC title

• C12Q2563/159

  Microreactors, e.g. emulsion PCR or sequencing, droplet PCR, microcapsules, i.e. non-liquid containers with a range of different permeability's for different reaction components · CPC title