# US 11,994,520 B2 · Developing lateral flow immunochromatography (LFIA) peptide-based test strips for rapid detection of antigens and antibodies against specific antigens > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 11,994,520 B2 - Document ID: US11994520B2 - Status: Granted - Kind: B2 - Application number: 18/038,107 - Filed: 2021-11-22 - Published: 2023-12-21 - Granted: 2024-05-28 - Earliest claimed: 2020-11-23 - Assignees: King Abdullah University of Science and Technology - Applicants: King Abdullah University of Science and Technology - Inventors: Charlotte A. E. Hauser; Panagiotis Bilalis; Dana Alhattab; Hepi Hari Susapto; Manola Moretti; Salwa Alshehri; Ali Aldoukhi; Hamed Albalawi; Hattan Boshah; Abdulelah Alrashoudi; Alexander Valle Perez; Rosario Perez Pedroza - Web page: https://patentgrep.com/patent/US11994520B2 ## CPC - G01N 33/56983: {Viruses} - C07K 14/005: from viruses - G01N 33/54388: {based on lateral flow} - G01N 33/558: using diffusion or migration of antigen or antibody {(immunochromatographic test strips G01N33/54387)} - C07K 2319/00: Fusion polypeptide - C07K 2319/40: containing a tag for immunodetection, or an epitope for immunisation - C12N 2770/20022: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes - G01N 2333/165: Coronaviridae, e.g. avian infectious bronchitis virus - G01N 2469/20: Detection of antibodies in sample from host which are directed against antigens from microorganisms - 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) - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) ## Abstract The present disclosure relates a method of fabricating a literal flow immunoassay (LFIA) for the diagnosis of diseases, including COVID-19. The present disclosure further relates to a fusion-epitopes peptide that can be used in the LFIA test to improve sensitivity, specificity and accuracy of the test. ## Claims ### Claim 1 A method of fabricating a lateral flow immunoassay (LFIA) comprising: synthesizing and screening at least one peptide antigen; conjugating the peptide antigen with at least one tag; printing a test strip; determining the minimum thickness of a test cassette; and assembling the LFIA, wherein the peptide antigen binds specifically to at least one analyst, wherein at least one control line and at least one test line are printed on the test strip, and wherein the material in the test line binds the complex formed by the analyst and the peptide antigen conjugated with the tag, wherein the epitopes are at least one selected from the group consisting of peptides with amino acid sequences provided in SEQ ID Nos. 1-10 and 10-19. ### Claim 2 (depends on 1) The method of claim 1, wherein the at least one peptide antigen is at least one fusion-epitopes peptide. ### Claim 3 (depends on 2) The method of claim 2, wherein the epitopes are whole or part of at least one antigenic protein of Acute Respiratory Syndrome Corona Virus 2 (SARS-CoV-2). ### Claim 4 (depends on 2) The method of claim 2, wherein the at least one fusion-epitopes peptide is obtained by fusing the peptide provided in SEQ ID No. 5 and at least one peptide provided in the group consisting SEQ ID Nos. 10, 13 and 19 or by fusing the peptide provided in SEQ ID No. 7 and the peptide provided in SEQ ID No. 8. ### Claim 5 (depends on 2) The method of claim 2, wherein the at least one fusion-epitopes peptide is provided in SEQ ID No. 11. ### Claim 6 (depends on 1) The method of claim 1, wherein the at least one analyst is at least one antibody. ### Claim 7 (depends on 1) The method of claim 1, wherein the at least one analyst is at least one anti-SARS-CoV-2 antibody. ### Claim 8 (depends on 1) The method of claim 1, wherein the minimum thickness of the test cassette is determined by a finite element analysis (FEA) simulation. ### Claim 9 (depends on 1) The method of claim 1, wherein the test cassette with the minimum thickness uses the least amount of materials to fabricate, when the cassettes does not break under handling forces of 5 N. ### Claim 10 (depends on 1) The method of claim 1, wherein the test cassette is printed. ### Claim 11 (depends on 1) The method of claim 1, wherein the at least one tag is at least one selected from the group consisting of colored nano-particle, silver nano-particle, carbon nano-particle, magnetic nano-particle, or quantum dot. ### Claim 12 (depends on 1) The method of claim 1, wherein the at least one tag is at least one selected from a gold nano-particle (AuNP) or a iron oxide nano-particle (Fe2O3). ### Claim 13 (depends on 1) The method of claim 1, further comprising: adding at least one agent binds to the control line but not the test line to the at least one tag conjugated peptide antigen. ### Claim 14 (depends on 1) The method of claim 1, wherein the at least one peptide antigen detects the at least one analyst with sensitivity of at least 85%, a specificity of at least 95%, and an accuracy of at least 85%. ### Claim 15 (depends on 1) The method of claim 1, wherein the at least one peptide antigen has at least 20 interactions with the at least one analyst and is determined using docking simulation. ### Claim 16 A fusion-peptide for immunoassay comprising at least two epitopes of SARS-CoV-2, wherein the epitopes are linked by peptide bond, wherein the epitopes are at least one selected from the group consisting of peptides with amino acid sequences provided in SEQ ID Nos. 1-10 and 10-19. ### Claim 17 (depends on 16) The fusion-peptide of claim 16, wherein the epitopes are different. ### Claim 18 (depends on 16) The fusion-peptide as in claim 16, wherein the fusion-epitopes peptide is obtained by fusing the peptide provided in SEQ ID No. 5 and at least one peptide provided in the group consisting SEQ ID Nos. 10, 13 and 19 or by fusing the peptide provided in SEQ ID No. 7 and the peptide provided in SEQ ID No. 8. ### Claim 19 (depends on 16) The fusion-peptide as in claim 16, wherein the fusion-epitopes peptide is provided in SEQ ID No. 11.