# US 12,602,559 B2 · Anti-counterfeit polynucleotide taggants

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- Number: US 12,602,559 B2
- Document ID: US12602559B2
- Status: Granted
- Kind: B2
- Application number: 19/184,702
- Filed: 2025-04-21
- Published: 2025-08-21
- Granted: 2026-04-14
- Earliest claimed: 2023-04-17
- Assignees: Microsoft Technology Licensing, LLC
- Applicants: Microsoft Technology Licensing, LLC
- Inventors: Yuan-Jyue Chen; Karin Strauss; Christian Paquin; Alexander Steven Crown; Sergey Yekhanin
- Web page: https://patentgrep.com/patent/US12602559B2

## CPC

- C12Q 1/6834: Enzymatic or biochemical coupling of nucleic acids to a solid phase
- G06K 7/1413: {1D bar codes}
- C12Q 1/6823: Release of bound markers
- G16B 50/30: Data warehousing; Computing architectures
- 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
- G06K: GRAPHICAL DATA READING (image or video recognition or understanding G06V); PRESENTATION OF DATA; RECORD CARRIERS; HANDLING RECORD CARRIERS
- G16B: BIOINFORMATICS, i.e. INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR GENETIC OR PROTEIN-RELATED DATA PROCESSING IN COMPUTATIONAL MOLECULAR BIOLOGY

## Abstract

Molecular anti-counterfeiting taggants are made from a plurality of synthetic polynucleotides that collectively encode a bit sequence using the sequences and hybridization states of the polynucleotides. The polynucleotide taggant is placed on an item as a molecular identifier of authenticity. The bit sequence encoded by the polynucleotide taggant is read out using a substrate which has bound polynucleotides complexes that hybridize with the synthetic polynucleotides in the polynucleotide taggant. A detectable signal is present where hybridization occurs. To prevent a bad actor from reverse engineering and creating a copy of the polynucleotide taggant using the results of hybridization to the substrate, multiple versions of the substrate are created. Each version hybridizes to different subsets of the synthetic polynucleotides in the polynucleotide taggant. A detectable pattern on the substrate that is present when exposed to the polynucleotide taggant is used for validating authenticity of the item.

## Claims

### Claim 1

A method of manufacturing a plurality of substrates to use for authenticating an item comprising:
generating on each of the plurality of substrates a plurality of physically separated assay locations that each comprise bound polynucleotide complexes that comprise a bound strand configured to attach to one of the assay locations and a reporter strand configured to hybridize to the bound strand, wherein one of the bound strand or the reporter strand includes a toehold domain that does not hybridize to the other strand and wherein each of the bound polynucleotide complexes includes a unique barcode domain indicating a specific bit position in a bit sequence; and
binding individual ones of the bound polynucleotide complexes to assay locations on the plurality of substrates such that at each individual assay location all the bound polynucleotide complexes have the same unique barcode domain and that a pattern of which unique barcode domains are bound at the assay locations is different for each substrate in the plurality of substrates.

### Claim 2 (depends on 1)

The method of claim 1, wherein the substrates are nitrocellulose membranes and generating a plurality of physically separated assay locations comprises depositing a pattern of hydrophobic barriers.

### Claim 3 (depends on 2)

The method of claim 2, wherein the hydrophobic barriers are formed of wax and generating the plurality of physically separated assay locations comprises depositing the wax with a wax-based printer.

### Claim 4 (depends on 1)

The method of claim 1, wherein the substrates are eggshell membranes (ESM), nylon membranes, nanofibers, or chitosan-modified membranes.

### Claim 5 (depends on 1)

The method of claim 1, wherein the plurality of physically separated assay locations includes at least nine separate assay locations on each substrate.

### Claim 6 (depends on 1)

The method of claim 1, further comprising synthesizing the bound polynucleotide complexes, wherein synthesizing the bound polynucleotide complexes comprises synthesizing each bound strand and each reporter strand followed by pre-hybridizing bound strands and reporter strands having the same unique barcode domain.

### Claim 7 (depends on 6)

The method of claim 6, wherein synthesizing the reporter strand comprises attaching a reporter molecule to the reporter strand.

### Claim 8 (depends on 7)

The method of claim 7, wherein the reporter molecule comprises a fluorophores, a dye, a magnetized material, or a radioactive material.

### Claim 9 (depends on 1)

The method of claim 1, further comprising, placing a validation code on each of the plurality of substrates.

### Claim 10 (depends on 1)

The method of claim 1, further comprising, incubating one of the substrates with a solution that contains a polynucleotide taggant, wherein the polynucleotide taggant comprises a plurality of synthetic polynucleotides that collectively encode a bit sequence using sequences of the synthetic polynucleotides and hybridization states of the synthetic polynucleotides, wherein the one of the substrates detects only a subset of bits in the bit sequence encoded by the polynucleotide taggant.

### Claim 11

A method of manufacturing a plurality of substrates to use for authenticating an item comprising:
generating on each of the plurality of substrates a plurality of physically separated assay locations;
synthesizing a plurality of bound polynucleotide complexes that each comprise a bound strand configured to attach to one of the assay locations and a reporter strand configured to hybridize to the bound strand, wherein one of the bound strand or the reporter strand includes a toehold domain that does not hybridize to the other strand and wherein each of the bound polynucleotide complexes includes a unique barcode domain indicating a specific bit position in a bit sequence; and
binding individual ones of the bound polynucleotide complexes to assay locations on the plurality of substrates such that at each individual assay location all the bound polynucleotide complexes have the same unique barcode domain and that a pattern of which unique barcode domains are bound at the assay locations is different for each substrate in the plurality of substrates.

### Claim 12 (depends on 11)

The method of claim 11, wherein the substrates are nitrocellulose membranes and generating a plurality of physically separated assay locations comprises depositing a pattern of hydrophobic barriers.

### Claim 13 (depends on 12)

The method of claim 12, wherein the hydrophobic barriers are formed of wax and generating the plurality of physically separated assay locations comprises depositing the wax with a wax-based printer.

### Claim 14 (depends on 11)

The method of claim 11, wherein the substrates are eggshell membranes (ESM), nylon membranes, nanofibers, or chitosan-modified membranes.

### Claim 15 (depends on 11)

The method of claim 11, wherein the plurality of physically separated assay locations includes at least nine separate assay locations on each substrate.

### Claim 16 (depends on 11)

The method of claim 11, wherein synthesizing the plurality of bound polynucleotide complexes comprises synthesizing each bound strand and each reporter strand followed by pre-hybridizing bound strands and reporter strands having the same unique barcode domain.

### Claim 17 (depends on 16)

The method of claim 16, wherein synthesizing the reporter strand comprises attaching a reporter molecule to the reporter strand.

### Claim 18 (depends on 17)

The method of claim 17, wherein the reporter molecule comprises a fluorophores, a dye, a magnetized material, or a radioactive material.

### Claim 19 (depends on 11)

The method of claim 11, further comprising, placing a validation code on each of the plurality of substrates.

### Claim 20 (depends on 11)

The method of claim 11, further comprising, incubating one of the substrates with a solution that contains a polynucleotide taggant, wherein the polynucleotide taggant comprises a plurality of synthetic polynucleotides that collectively encode a bit sequence using sequences of the synthetic polynucleotides and hybridization states of the synthetic polynucleotides, wherein the one of the substrates detects only a subset of bits in the bit sequence encoded by the polynucleotide taggant.