US 12,436,148 B2
Direct immunoassay measurement of autoantibodies
James Freeman, Sandy Hook, CT (US); Mariah Arcuri, Norwood, NJ (US); Cheryl Krumm, Yonkers, NY (US); and Eddine Merabet, White Plains, NY (US)
Assigned to Siemens Healthcare Diagnostics Inc., Tarrytown, NY (US)
Appl. No. 17/250,151
Filed by Siemens Healthcare Diagnostics Inc., Tarrytown, NY (US)
PCT Filed Jul. 1, 2019, PCT No. PCT/US2019/040167
§ 371(c)(1), (2) Date Dec. 7, 2020,
PCT Pub. No. WO2020/010009, PCT Pub. Date Jan. 9, 2020.
Claims priority of provisional application 62/693,439, filed on Jul. 2, 2018.
Prior Publication US 2021/0247392 A1, Aug. 12, 2021
Int. Cl. G01N 33/564 (2006.01); G01N 33/543 (2006.01); G01N 33/78 (2006.01)
CPC G01N 33/564 (2013.01) [G01N 33/54353 (2013.01); G01N 33/78 (2013.01); G01N 2800/24 (2013.01)] 9 Claims
 
1. A method of diagnosing an autoimmune thyroid disease in a subject, the method comprising:
a) diluting a biological sample from the subject to form a diluted biological sample and incubating the diluted biological sample from the subject with:
a solid support having an unlabeled thyroglobulin bound thereto, wherein the unlabeled thyroglobulin is specifically recognized by an anti-thyroglobulin autoantibody from the subject, wherein the solid support comprises a magnetic bead or a magnetic particle; and
a chemiluminescent-labeled thyroglobulin comprising a chemiluminescent label bound to a thyroglobulin in at least a 10 times molar excess to the thyroglobulin, wherein the chemiluminescent-labeled thyroglobulin does not bind to the solid support and is specifically recognized by the anti-thyroglobulin autoantibody,
wherein, in the presence of the anti-thyroglobulin autoantibody, a solid support/chemiluminescent-labeled thyroglobulin complex is formed;
b) separating with a magnet the solid support/chemiluminescent-labeled thyroglobulin complex from the chemiluminescent-labeled thyroglobulin that is not bound by the anti-thyroglobulin autoantibody; and
c) detecting the chemiluminescence level of the solid support/chemiluminescent-labeled thyroglobulin complex, wherein the solid support/chemiluminescent-labeled thyroglobulin complex comprises a chemiluminescent label comprising acridinium ester or an analog thereof,
thereby diagnosing the subject with the autoimmune thyroid disease.