# US 11,029,319 B2 · Biosensor and application of the same > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 11,029,319 B2 - Document ID: US11029319B2 - Status: Granted - Kind: B2 - Application number: 16/157,094 - Filed: 2018-10-11 - Published: 2020-04-16 - Granted: 2021-06-08 - Earliest claimed: 2018-10-11 - Assignees: National Cheng Kung University - Applicants: National Cheng Kung University - Inventors: Han-Sheng Chuang; Ching-Chuen Chen; Yu-Ting Tseng - Web page: https://patentgrep.com/patent/US11029319B2 ## CPC - G01N 33/6893: {related to diseases not provided for elsewhere} - A61B 5/14507: {specially adapted for measuring characteristics of body fluids other than blood (devices for taking samples of body liquids A61B10/0045)} - A61B 5/1455: using optical sensors, e.g. spectral photometrical oximeters - A61B 5/6821: {Eye} - A61B 10/0045: {Devices for taking samples of body liquids} - G01N 21/64: Fluorescence; Phosphorescence - G01N 21/645: {Specially adapted constructive features of fluorimeters} - G01N 21/774: {the reagent being on a grating or periodic structure} - G02C 7/04: Contact lenses for the eyes (disinfection or sterilisation of contact lenses A61L12/00) - G02C 11/10: {Electronic devices other than hearing aids} - A61B 2010/0067: {Tear or lachrymal fluid} - B82Y 30/00: Nanotechnology for materials or surface science, e.g. nanocomposites - G01N 2021/6463: {Optics} - G01N 2333/47: Assays involving proteins of known structure or function as defined in the subgroups - G01N 2800/164: Retinal disorders, e.g. retinopathy - 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) - A61B: DIAGNOSIS; SURGERY; IDENTIFICATION (usefulness limited to only animals A61D) - G02C: SPECTACLES; SUNGLASSES OR GOGGLES INSOFAR AS THEY HAVE THE SAME FEATURES AS SPECTACLES; CONTACT LENSES (trial frames for testing the eyes A61B3/04; goggles or eyeshields not having the same features as spectacles A61F9/00) - B82Y: SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES ## Abstract The present invention provides a biosensor and an application of the same. The biosensor includes a substrate, a first polymer layer and a second polymer layer. The first polymer layer includes composite antibodies, each of which includes a first antibody and a labelling molecule. The second polymer layer has an inverse opal photonic crystal structure where gold nanoparticles and second antibodies are distributed. At least one of the composite antibodies, an antigen and at least one of the second antibodies forms a complex in the second polymer layer, and an antigen concentration is obtained by a fluorescence intensity, a degree of red-shift or a change in a visual color of the biosensor. ## Claims ### Claim 1 A biosensor, comprising: a substrate, wherein the substrate comprises a first region and a second region adjoined to the first region, the second region is located on one side of the first region, a center of the substrate is concentrically surrounded by the first region and the second region in order from outside to inside, a bottom of the first region has a recessed cross section having a first depth, a bottom of the second region has a recessed cross section having an asymmetric U-shape and a second depth, the second depth is greater than the first depth, and a slope of a side wall of the recessed cross section of the second region near the center is greater than a slope of another other side wall of the recessed cross section of the second region near the first region; a first polymer layer, disposed in the first region, wherein a plurality of composite antibodies are distributed in the first polymer layer, and each of the composite antibodies comprises a labelling molecule and a first antibody connected to the labelling molecule; and a second polymer layer, disposed in the second region, wherein the second polymer layer has an inverse opal photonic crystal structure, the inverse opal photonic crystal structure comprises a plurality of holes, a plurality of gold nanoparticles and a plurality of second antibodies are disposed on a wall of each of the holes, and the first antibody and the second antibodies recognize the same antigen. ### Claim 2 (depends on 1) The biosensor of claim 1, wherein the second region is located over the first region. ### Claim 3 (depends on 1) The biosensor of claim 1, wherein the inverse opal photonic crystal structure is an inverted structure of a face-centered cubic structure of nanobeads, each of the nanobeads has a particle size in a range from 100 nm to 1000 nm, and the gold nanoparticles are distributed on a surface of each of the nanobeads. ### Claim 4 (depends on 1) The biosensor of claim 1, wherein the labelling molecule comprises a fluorescent molecule, and a particle size of each of the gold nanoparticles is in a range from 5 nm to 80 nm. ### Claim 5 (depends on 1) The biosensor of claim 1, further comprising a third polymer layer over the first polymer layer. ### Claim 6 (depends on 1) The biosensor of claim 1, wherein the biosensor is a contact lens, the center of the substrate is an optical zone of the contact lens, the first region and the second region are located in a non-optical zone of the contact lens, at least the second polymer layer is exposed on one surface of the contact lens, and opposes a surface that is in direct contact with an eye. ### Claim 7 (depends on 1) A method of detecting an antigen concentration, comprising: providing a biosensor described in claim 1, wherein a substrate of the biosensor comprises a first region and a second region connected to the first region, and the second region is located on one side of the first region; flowing a biological liquid sample containing an antigen from the second region to the first region of the biosensor, thereby releasing at least one of a plurality of composite antibodies comprising a first antibody and a labelling molecule, and reacting the at least one of the composite antibodies for a period of time with the antigen and at least one of a plurality of second antibodies to form a complex in the second region; detecting an optical property of the complex in the second region of the biosensor by using a light source having a specific wavelength, wherein the optical property comprises a fluorescence intensity or a visual color; and obtaining an antigen concentration according to the optical property. ### Claim 8 (depends on 7) The method of claim 7, wherein the specific wavelength is in a range from 200 nm to 700 nm, and the fluorescence intensity is increased by at least two times based on an inverse opal photonic crystal structure in the second region.