	US 11,953,503 B2
	Method for quantitatively determining a therapeutic TNF-alpha inhibitor
Harald Althaus, Wetter (DE); and Thorsten Keller, Rauschenberg (DE)
Assigned to Siemens Healthcare Diagnostics Products GmbH, Marburg (DE)
Appl. No. 16/962,566
Filed by Siemens Healthcare Diagnostics Products GmbH, Marburg (DE)
PCT Filed Jan. 14, 2019, PCT No. PCT/EP2019/050760 § 371(c)(1), (2) Date Jul. 16, 2020, PCT Pub. No. WO2019/141614, PCT Pub. Date Jul. 25, 2019.
Claims priority of application No. 18152009 (EP), filed on Jan. 17, 2018.
Prior Publication US 2020/0400681 A1, Dec. 24, 2020
Int. Cl. G01N 33/68 (2006.01); G01N 33/543 (2006.01)

CPC G01N 33/6863 (2013.01) [G01N 33/5434 (2013.01); G01N 2333/525 (2013.01)]

9 Claims

1. A method for quantitatively determining a therapeutic tumor necrosis factor (TNF)-alpha inhibitor in a sample, the method comprising the steps of:

a) contacting the sample

with isolated, free TNF-alpha protein to provide a reaction mixture;

b) incubating the reaction mixture to allow binding of the isolated, free TNF-alpha protein to therapeutic TNF-alpha inhibitor in the sample;

c) contacting the incubated reaction mixture with a first TNF-alpha binding partner to allow formation of a complex of isolated, free TNF-alpha protein remaining after step b) with the first TNF-alpha binding partner;

d) determining the amount of the complex composed of TNF-alpha protein and the first TNF-alpha binding partner; and

e) determining the amount of the therapeutic TNF-alpha inhibitor in the sample by comparing the amount of the complex composed of TNF-alpha protein and the first TNF-alpha binding partner determined in step d) with amounts of a complex composed of TNF-alpha protein and the first TNF-alpha binding partner in reaction mixtures containing samples having known concentrations of a TNF-alpha inhibitor.