# US 2026/0243760 A1 · Method and system for optically based immunoassays

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- Number: US 2026/0243760 A1
- Document ID: US20260243760A1
- Status: Published application
- Kind: A1
- Application number: 19/165,225
- Filed: 2024-03-20
- Published: 2026-08-20
- Earliest claimed: 2023-04-03
- Inventors: Martin Huber; Ossama Assad
- Web page: https://patentgrep.com/patent/US20260243760A1

## CPC

- G01N 33/54326: {Magnetic particles}
- G02B 21/16: adapted for ultraviolet illumination {; Fluorescence microscopes (G02B21/0076 takes precedence)}
- G01N 21/6428: {Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" (in vivo A61B5/00; immunoassay G01N33/53)}
- G01N 21/6452: {Individual samples arranged in a regular 2D-array, e.g. multiwell plates}
- G01N 21/648: {using evanescent coupling or surface plasmon coupling for the excitation of fluorescence}
- G01N 33/54386: {Analytical elements}
- G01N 33/54313: {the carrier being characterised by its particulate form}
- G01N 33/582: {with fluorescent label}
- G01N 2021/6419: {Excitation at two or more wavelengths}
- G01N 2021/6421: {Measuring at two or more wavelengths}
- G01N 2021/6439: {with indicators, stains, dyes, tags, labels, marks}
- G01N 2470/04: Sandwich assay format
- 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)
- G02B: OPTICAL ELEMENTS, SYSTEMS OR APPARATUS

## Abstract

The invention is directed to methods and systems for optically based immunoassays for one or more analytes, such as proteins, employing subwavelength hole arrays.

## Claims

### Claim 1

A method of determining the presence and amounts of one or more analytes in a sample, the method comprising:
providing a hole array comprising a solid phase membrane and an opaque layer co-extensive therewith and a plurality of holes there through, wherein (i) the opaque layer substantially prevents excitation light from passing through the hole array, and (ii) each hole has an exit and a diameter less than a wavelength of excitation light such that a signal generation region is formed adjacent to its exit whenever excitation light is incident to the opaque layer;
combining a sample with capture agents and associated detection agents to form one or more capture complexes each having one or more optical labels, wherein each capture agent and associated detection agents is specific for an analyte;
directing excitation light to the opaque layer of the hole array to produce signal generation regions at the exits of the holes;
translocating capture complexes through the hole array from a side opposite the opaque layer so that the one or more optical labels of each capture complex produce one or more optical signals as the capture complex passes through a signal generation region at the exits of the holes;
detecting for each hole the one or more optical signals of the translocated capture complexes;
determining the presence or absence and the quantities of each of the one or more analytes in the sample from the optical signals collected from the capture complexes.

### Claim 2 (depends on 1)

The method of claim 1 wherein said opaque layer is a metal layer.

### Claim 3 (depends on 2)

The method of claim 2 wherein said metal layer comprises an aluminum layer or a gold layer.

### Claim 4 (depends on 2)

The method of claim 2 wherein said directing and detecting are implemented with an epi-illumination system.

### Claim 5 (depends on 1)

The method of claim 1 wherein said capture agent comprises a particle having a diameter less than said diameter of said holes.

### Claim 6 (depends on 5)

The method of claim 5 wherein said particle is magnetic.

### Claim 7 (depends on 5)

The method of claim 5 wherein said particle is charged.

### Claim 8 (depends on 1)

The method of claim 1 wherein said optical labels are fluorescent labels.

### Claim 9 (depends on 1)

The method of claim 1 wherein each of said capture complexes comprises an analyte sandwiched between a capture antibody and a detection antibody.

### Claim 10 (depends on 9)

The method of claim 9 wherein said one or more analytes comprise one or more proteins.

### Claim 11 (depends on 1)

The method of claim 1 wherein each of said capture complexes comprises a capture antibody and an unlabeled analyte or a labeled analyte.

### Claim 12 (depends on 1)

The method of claim 1 wherein each of said capture agents comprise an allergen.

### Claim 13

A method of determining the presence and amounts of one or more analytes in a sample, the method comprising:
combining a sample with capture agents and associated detection agents to form one or more capture complexes each having one or more optical labels, wherein each capture agent and associated detection agents is specific for an analyte;
translocating capture complexes through a hole array illuminated with excitation light, wherein the hole array comprises a solid phase membrane and an opaque layer co-extensive therewith and a plurality of holes there through, such that (i) the opaque layer substantially prevents excitation light from passing through the hole array, and (ii) each hole has an exit and a diameter less than a wavelength of excitation light such that a signal generation region is formed adjacent to its exit;
detecting for each hole the one or more optical signals of the translocated capture complexes; and
determining the presence or absence and the quantities of each of the one or more analytes in the sample from the optical signals collected from the capture complexes.

### Claim 14 (depends on 13)

The method of claim 13 wherein said opaque layer is a metal layer.

### Claim 15 (depends on 13)

The method of claim 13 wherein each of said capture agent comprises a particle having a diameter less than said diameter of said holes.

### Claim 16 (depends on 15)

The method of claim 15 wherein ninety percent or more of said one or more optical signals indicate said particles comprise 0, 1 or 2 bound analytes

### Claim 17 (depends on 15)

The method of claim 15 wherein said particle is magnetic.

### Claim 18 (depends on 15)

The method of claim 15 wherein said particle is charged.

### Claim 19 (depends on 13)

The method of claim 13 wherein said optical labels are fluorescent labels.

### Claim 20 (depends on 13)

The method of claim 13 wherein each of said capture complexes comprises an analyte sandwiched between a capture antibody and a detection antibody.

### Claim 21 (depends on 20)

The method of claim 20 wherein said one or more analytes comprise one or more proteins.

### Claim 22 (depends on 13)

The method of claim 13 wherein each of said capture complexes comprises a capture antibody and an unlabeled analyte or a labeled analyte.

### Claim 23 (depends on 13)

The method of claim 13 wherein each of said capture agents comprise an allergen.

### Claim 24

A system for determining the presence and amounts of one or more analytes in a sample, the system comprising:
a hole array comprising a solid phase membrane and an opaque layer co-extensive therewith and a plurality of holes therethrough, such that (i) the opaque layer substantially prevents excitation light directed thereto from passing through the hole array, and (ii) each hole has an exit and a diameter less than a wavelength of excitation light such that a signal generation region is formed adjacent to its exit;
one or more beads each having a diameter less than the diameters of the holes of the hole array and each comprising a capture agent specific for an analyte, wherein the one or more beads form capture complexes in the presence of analytes and detection agents, such that the capture complexes generate one or more optical signals as such capture complexes translocate through the signal generation regions of the holes of the hole array, thereby generating optical signals which are indicative of the presence and amounts of the analytes in the sample.

### Claim 25 (depends on 24)

The system of claim 24 further comprising;
One or more light sources that direct one or more excitation beams to said opaque layer of said hole array;
a cis reservoir on a side of said hole array opposite of said opaque layer; and
a trans reservoir on a side of said hole array the same as the excitation beam, wherein said capture complexes are translocated from the cis reservoir to the trans reservoir through said signal generation regions of said holes of said hole array.

### Claim 26 (depends on 25)

The system of claim 25 further comprising one or more detectors for collecting said optical signals.