US 11,988,668 B2
Concentration-dependent self-interaction assay
Michael Marlow, New Rochelle, NY (US); Michael Sennett, Somerville, MA (US); and Michael Schneider, Tarrytown, NY (US)
Assigned to Regeneron Pharmaceuticals, Inc., Tarrytown, NY (US)
Filed by Regeneron Pharmaceuticals, Inc., Tarrytown, NY (US)
Filed on Jul. 20, 2022, as Appl. No. 17/868,983.
Application 17/868,983 is a division of application No. 16/326,238, granted, now 11,428,695, previously published as PCT/US2017/047630, filed on Aug. 18, 2017.
Claims priority of provisional application 62/376,788, filed on Aug. 18, 2016.
Prior Publication US 2022/0357335 A1, Nov. 10, 2022
Int. Cl. G01N 33/68 (2006.01); G01N 21/25 (2006.01); G01N 21/31 (2006.01); G01N 21/77 (2006.01); G01N 33/58 (2006.01)
CPC G01N 33/6845 (2013.01) [G01N 21/77 (2013.01); G01N 33/587 (2013.01)] 19 Claims
 
1. A method of making a low viscosity pharmaceutical formulation containing a protein, the method comprising:
a. determining the potential of the protein to self-associate, wherein the determining step comprises:
(i) combining the protein at a low concentration, a nanoparticle, and a buffered salt to form a sample;
(ii) exciting the sample with light;
(iii) measuring the light transmitted through the sample at multiple wavelengths ranging from 450 nm to about 750 nm;
(iv) calculating the absorbance intensity ratio of the sample, wherein the absorbance intensity ratio is the ratio of absorbance intensity at the maximum absorbance (λmax) to the initial absorbance at 450 nm; and
(v) measuring the absorbance intensity ratio, wherein an absorbance ratio above 1.7 (Apeak/(Ainit) indicates that the protein retains 90% of its native structure at a high concentration;
b. combining a viscosity-reducing excipient with the protein;
c. determining the potential of the protein to self-associate in the presence of the viscosity-reducing excipient; and
d. formulating the protein with the viscosity-reducing excipient at a level that reduces the viscosity of the protein solution by at least 50% than without the viscosity-reducing excipient.