# US 2018/0024137 A1 · Methods for comparing a structure of a first biomolecule and a second biomolecule > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2018/0024137 A1 - Document ID: US20180024137A1 - Status: Published application - Kind: A1 - Application number: 15/720,755 - Filed: 2017-09-29 - Published: 2018-01-25 - Earliest claimed: 2013-06-04 - Inventors: Sarfaraz K. Niazi - Web page: https://patentgrep.com/patent/US20180024137A1 ## CPC - G01N 33/6803: {General methods of protein analysis not limited to specific proteins or families of proteins} - G01N 21/6486: {Measuring fluorescence of biological material, e.g. DNA, RNA, cells (G01N21/6428 takes precedence)} - 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) ## Abstract The present disclosure provides methods to assess structural similarity of a first biomolecule and a second biomolecule by detecting one or more responses of the first and second biomolecule to thermodynamic stress conditions induced by osmotic and dielectric changes including, detecting a shift in fluorescence emission and/or a change in the intensity of the emission. ## Claims ### Claim at position 1 1-19. (canceled) ### Claim at position 2 20: A method of comparing structural similarity of a first protein to a second protein that each have a fluorescence emission property, the method comprising: a) providing a first solution comprising a component at a first concentration and the first protein; b) altering the first concentration of the component in the first solution to a second concentration to form a second solution, wherein the second concentration changes the fluorescence emission property of the first protein; c) measuring the fluorescence emission property of the first protein in the second solution; d) comparing the fluorescence emission property of the first protein in the second solution to the fluorescence emission property of the second protein in a third solution comprising the component at the second concentration, wherein the second protein is a biosimilar of the first protein. ### Claim at position 3 21: A method of comparing structural similarity of a first protein to a second protein that each have a fluorescence emission property, the method comprising: a) providing a first solution comprising a component at a first concentration and the first protein; b) altering the first concentration of the component to a second concentration to form a second solution, wherein the second concentration changes the fluorescence emission property of the first protein; c) measuring the change in the fluorescence emission property of the first protein in the first solution and second solution; d) providing a third solution comprising the component at the first concentration and the second protein, wherein the second protein is a biosimilar of the first protein; e) altering the concentration of the component of the third solution to the second concentration to form a fourth solution; f) measuring a change in the fluorescence emission property of the second protein in the third solution and fourth solution; g) comparing the change in c) and the change in f). ### Claim at position 4 (depends on 20, 21) 22: The method of claim 20 or claim 21, wherein the first protein and second protein are manufactured by different processes. ### Claim at position 5 (depends on 22) 23: The method of claim 22, wherein the first protein and second protein are manufactured using different host cells. ### Claim at position 6 (depends on 22) 24: The method of claim 22, wherein the first protein and second protein are manufactured using different fermentation processes. ### Claim at position 7 (depends on 22) 25: The method of claim 22, wherein the first protein and second protein are manufactured using different purification processes. ### Claim at position 8 (depends on 20, 21) 26: The method according to claim 20 or claim 21, wherein the component comprises one or more osmolytes. ### Claim at position 9 (depends on 26) 27: The method according to claim 26, wherein the one or more osmolytes are selected from the group consisting of a glycerol, a polyethylene glycol, a buffer, a salt, urea, a non-ionic detergent, an ionic detergent, a non-ionic surfactant, an ionic surfactant, an acid, a hydrophobic molecule, a natural osmolyte, and combinations thereof. ### Claim at position 10 (depends on 26) 28: The method according to claim 26, wherein the osmolyte is acetate buffer. ### Claim at position 11 (depends on 27) 29: The method according to claim 27, wherein the natural osmolyte is selected from the group consisting of trimethylamine N-oxide (TMAO), dimethylsulfoniopropionate, trimethylglycine, sarcosine, betaine, glycerophosphorylcholine, myoinositol, and taurine. ### Claim at position 12 (depends on 20, 21) 30: The method according to claim 20 or 21, wherein the component comprises polysorbate. ### Claim at position 13 (depends on 20, 21) 31: The method according to claim 20 or 21, wherein the fluorescence emission property is fluorescence emission wavelength. ### Claim at position 14 (depends on 20, 21) 32: The method according to claim 20 or 21, wherein the fluorescence emission property is fluorescence emission intensity. ### Claim at position 15 (depends on 20, 21) 33: The method according to claim 20 or 21, wherein the fluorescence emission property is measured using an excitation wavelength between 150 and 300 nm. ### Claim at position 16 (depends on 31) 34: The method according to claim 31, wherein the excitation wavelength is 257 nm. ### Claim at position 17 (depends on 31) 35: The method according to claim 31, wherein the excitation wavelength is 274 nm. ### Claim at position 18 (depends on 31) 36: The method according to claim 31, wherein the excitation wavelength is 280 nm. ### Claim at position 19 (depends on 20, 21) 37: The method according to claim 20 or 21, wherein the first protein and second protein each comprise a fluorescent active amino acid residue selected from the group consisting of tyrosine, tryptophan, and phenylalanine. ### Claim at position 20 (depends on 20, 21) 38: The method according to claim 20 or 21, wherein the first protein and second protein are selected from the group consisting of: antibodies, antibody fragments, vaccines, therapeutic proteins, enzymes, protein digests, and denatured proteins. ### Claim at position 21 (depends on 37) 39: The method according to claim 37, wherein the first protein and second protein are therapeutic proteins. ### Claim at position 22 (depends on 37) 40: The method according to claim 37, wherein the first protein and second protein are antibodies.