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US 2018/0024137 A1Published application
Methods for comparing a structure of a first biomolecule and a second biomolecule
OwnerSarfaraz K. Niaziinventor
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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.
Claims22
1-19. (canceled)
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.
2 more elements
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).
5 more elements
22:
- The method of claim 20 or claim 21, wherein the first protein and second protein are manufactured by different processes.
23:
- The method of claim 22, wherein the first protein and second protein are manufactured using different host cells.
24:
- The method of claim 22, wherein the first protein and second protein are manufactured using different fermentation processes.
25:
- The method of claim 22, wherein the first protein and second protein are manufactured using different purification processes.
26:
- The method according to claim 20 or claim 21, wherein the component comprises one or more osmolytes.
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.
28:
- The method according to claim 26, wherein the osmolyte is acetate buffer.
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.
30:
- The method according to claim 20 or 21, wherein the component comprises polysorbate.
31:
- The method according to claim 20 or 21, wherein the fluorescence emission property is fluorescence emission wavelength.
32:
- The method according to claim 20 or 21, wherein the fluorescence emission property is fluorescence emission intensity.
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.
34:
- The method according to claim 31, wherein the excitation wavelength is 257 nm.
35:
- The method according to claim 31, wherein the excitation wavelength is 274 nm.
36:
- The method according to claim 31, wherein the excitation wavelength is 280 nm.
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.
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.
39:
- The method according to claim 37, wherein the first protein and second protein are therapeutic proteins.
40:
- The method according to claim 37, wherein the first protein and second protein are antibodies.