US 12,241,849 B2
Methods of determining protein or peptide concentration and uses thereof
Scott Allan Bradley, Brownsburg, IN (US); William F. Weiss, IV, Indianapolis, IN (US); and Wesley Clinton Jackson, Jr., Indianapolis, IN (US)
Assigned to Eli Lilly and Company, Indianapolis, IN (US)
Filed by Eli Lilly and Company, Indianapolis, IN (US)
Filed on Jan. 22, 2021, as Appl. No. 17/155,594.
Application 17/155,594 is a continuation of application No. PCT/US2019/046437, filed on Aug. 14, 2019.
Claims priority of provisional application 62/727,708, filed on Sep. 6, 2018.
Claims priority of provisional application 62/720,607, filed on Aug. 21, 2018.
Prior Publication US 2021/0215625 A1, Jul. 15, 2021
Int. Cl. G01N 24/08 (2006.01); G01N 33/487 (2006.01); G01N 33/68 (2006.01); G01R 33/46 (2006.01); G01R 33/465 (2006.01)
CPC G01N 24/08 (2013.01) [G01N 24/085 (2013.01); G01N 24/088 (2013.01); G01N 33/487 (2013.01); G01N 33/6803 (2013.01); G01R 33/4616 (2013.01); G01R 33/465 (2013.01); G01N 2496/00 (2013.01)] 19 Claims
OG exemplary drawing
 
1. A method of determining absolute protein concentration of a protein in a drug substance or a drug product solution, wherein the drug substance or drug product contains a single protein, comprising the steps of:
a. denaturing the protein, wherein the denaturing causes the protein to become unfolded and lose a majority of its secondary, tertiary, and quaternary structural features;
b. performing qNMR spectroscopy on the denatured protein, wherein said step of performing qNMR spectroscopy includes use of a diffusion filter; and
c. calculating the absolute protein concentration of the protein using a reference standard and a referencing technique.