# US 2026/0086090 A1 · Ultra-sensitive analyte detection and quantification using catch and release with proximity detection

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- Number: US 2026/0086090 A1
- Document ID: US20260086090A1
- Status: Published application
- Kind: A1
- Application number: 19/112,170
- Filed: 2023-09-14
- Published: 2026-03-26
- Earliest claimed: 2022-09-15
- Assignees: Children's Medical Center Corporation
- Applicants: The Children's Medical Center Corporation
- Inventors: Clinton H. Hansen; Wesley Philip Wong
- Web page: https://patentgrep.com/patent/US20260086090A1

## CPC

- G01N 33/542: with steric inhibition or signal modification, e.g. fluorescent quenching
- G01N 33/56983: {Viruses}
- C12Q 1/6804: Nucleic acid analysis using immunogens (immunoassay G01N33/53)
- G01N 33/5308: {for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites}
- G01N 33/54306: {Solid-phase reaction mechanisms}
- G01N 2333/165: Coronaviridae, e.g. avian infectious bronchitis virus
- G01N 2458/10: Oligonucleotides as tagging agents for labelling antibodies
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES

## Abstract

Provided herein is a highly sensitive analyte detection method that involves catch and release of a target analyte coupled with a proximity assay. Target analyte is captured using a capture binding partner and is also bound by detection binding partners thereby forming an analyte-biding partner complex. The complex is immobilized, and unbound reactants are physically removed. Thereafter the complex is released and a proximity assay is performed on the complex in order to detects its presence.

## Claims

### Claim 1

A method of detecting a target analyte, comprising
combining a capture binding partner and a detection binding partner with a sample, under conditions sufficient for the capture binding partner and detection binding partner to bind to a target analyte present in the sample, thereby forming an analyte-binding partner complex
immobilizing the analyte-binding partner complex to a solid support,
removing unbound capture binding partner and detection binding partner,
releasing the analyte-binding partner complex,
transferring the released analyte-binding partner complex to a second container
performing a proximity assay on the released analyte-binding partner complex in solution, thereby detecting the presence of the analyte-binding partner complex.

### Claim 2

A method of detecting a target analyte, comprising
forming an analyte-binding partner complex comprising a target analyte bound simultaneously to a capture binding partner and two or more detection binding partners, wherein the capture binding partner is immobilized to a solid support before or after binding to the target analyte,
separating the analyte-binding partner complex from unbound capture binding partner and/or unbound detection binding partner,
releasing immobilized analyte-binding partner complex,
transferring the released analyte-binding partner complex to a second container
performing a surface-free proximity assay on the released analyte-binding partner complex, thereby detecting the presence of the analyte-binding partner complex.

### Claim 3 (depends on 1, 2)

The method of claim 1 or 2, wherein the target analyte is a protein or a peptide.

### Claim 4 (depends on 1, 2)

The method of claim 1 or 2, wherein the capture binding partner is amino acid or nucleic acid in nature.

### Claim 5 (depends on 1, 2)

The method of claim 1 or 2, wherein the capture binding partner is an antibody or antigen-binding antibody fragment, a nanobody, a single chain antibody, or an aptamer.

### Claim 6 (depends on 1, 2)

The method of claim 1 or 2, wherein the detection binding partner comprises an analyte-specific binding partner conjugated to a detection moiety.

### Claim 7 (depends on 6)

The method of claim 6, wherein the analyte-specific binding partner is amino acid or nucleic acid in nature.

### Claim 8 (depends on 6)

The method of claim 6, wherein the analyte-specific binding partner is an antibody or antigen-binding antibody fragment, a nanobody, a single chain antibody, or an aptamer.

### Claim 9 (depends on 6)

The method of claim 6, wherein the detection moiety is an enzyme substrate, an enzyme, FRET donor or a FRET acceptor.

### Claim 10 (depends on 6)

The method of claim 6, wherein the detection moiety is an oligonucleotide.

### Claim 11 (depends on 1, 2)

The method of claim 1 or 2, wherein the sample is a bodily sample, optionally a saliva sample, a sputum sample, or a nose or nasopharyngeal swab sample.

### Claim 12 (depends on 1, 2)

The method of claim 1 or 2, wherein the target analyte is produced by a pathogen, optionally a virus, a bacterium, or a fungus.

### Claim 13 (depends on 1, 2)

The method of claim 1 or 2, wherein the target analyte is SARS-COV-2 spike protein.

### Claim 14 (depends on 1, 2)

The method of claim 1 or 2, wherein the target analyte is present in the sample at a concentration of about or at least 5 attoMolar.

### Claim 15 (depends on 1, 2)

The method of claim 1 or 2, wherein the solid support is a surface of a well or tube or bead.

### Claim 16 (depends on 1, 2)

The method of claim 1 or 2, wherein the capture binding partner is conjugated to a first oligonucleotide, the solid support is conjugated to a second oligonucleotide, the capture binding partner is immobilized to the solid support by hybridization of the first and second oligonucleotides to each other, and wherein the analyte-binding partner complex is released by nucleic acid strand displacement in the presence of a release oligonucleotide, optionally wherein the release oligonucleotide is complementary to the second oligonucleotide.

### Claim 17 (depends on 1, 2)

The method of claim 1 or 2, wherein the capture binding partner is immobilized to the solid support by a ultraviolet (UV) cleavable linker, and the analyte-binding partner complex is released by exposure to ultraviolet (UV) light.

### Claim 18 (depends on 1, 2)

The method of claim 1 or 2, wherein the capture binding partner is immobilized to the solid support by a nucleic acid linker, and the analyte-binding partner complex is released by contact with a restriction enzyme.

### Claim 19 (depends on 1, 2)

The method of claim 1 or 2, wherein during or following the release of the analyte-binding partner complex the solid support is exposed to a quenching agent to prevent re-association of the released analyte-binding partner complex.

### Claim 20 (depends on 1, 2)

The method of claim 1 or 2, wherein the detection binding partner is two detection binding partners, and each detection binding partner is conjugated to an oligonucleotide that is a substrate in a proximity assay.

### Claim 21 (depends on 1, 2)

The method of claim 1 or 2, wherein the detection binding partner is three detection binding partners, and each detection binding partner is conjugated to an oligonucleotide that is a substrate in a proximity assay.

### Claim 22 (depends on 1, 2)

The method of claim 1 or 2, wherein the proximity assay is a proximity ligation assay.

### Claim 23 (depends on 1, 2)

The method of claim 1 or 2, wherein the proximity assay is a proximity extension assay.