US 12,442,030 B2
Methods and systems for determining ADAMTS13 enzyme activity
Russell Philip Grant, Chapel Hill, NC (US); Christopher Michael Shuford, Mebane, NC (US); and Meghan Norris Bradley, Mebane, NC (US)
Assigned to Laboratory Corporation of America Holdings, Burlington, NC (US)
Filed by Laboratory Corporation of America Holdings, Burlington, NC (US)
Filed on Mar. 7, 2024, as Appl. No. 18/598,286.
Application 18/598,286 is a division of application No. 17/088,778, filed on Nov. 4, 2020, granted, now 11,959,124, issued on Apr. 16, 2024.
Application 17/088,778 is a continuation of application No. 15/730,165, filed on Oct. 11, 2017, granted, now 10,858,689, issued on Dec. 8, 2020.
Claims priority of provisional application 62/406,693, filed on Oct. 11, 2016.
Prior Publication US 2024/0271179 A1, Aug. 15, 2024
Int. Cl. C12Q 1/37 (2006.01); C12Q 1/00 (2006.01); G01N 33/68 (2006.01)
CPC C12Q 1/37 (2013.01) [C12Q 1/005 (2013.01); G01N 33/6851 (2013.01); G01N 2333/96486 (2013.01); G01N 2560/00 (2013.01); G01N 2800/226 (2013.01)] 19 Claims
 
1. A system for determining activity of enzyme disintegrin and metalloproteinase with a thrombospondin type 1 motif, member 13 (ADAMTS13) in a sample, the system comprising:
a station for incubating the sample with an exogenous peptide substrate for ADAMTS13 under conditions allowing for enzymatic cleavage of the exogenous peptide substrate by ADAMTS13 to generate an enzymatic cleavage product, wherein the exogenous peptide substrate consists of an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 3 and comprises an ADAMTS13 cleavage site;
a station for ionizing the enzymatic cleavage product to generate a multiply charged gas-phase ion of the enzymatic cleavage product; and,
a station for analyzing the multiply charged gas-phase ion by tandem mass spectrometry to determine presence and/or amount of the enzymatic cleavage product in the sample, wherein the presence or the amount of the enzymatic cleavage product is indicative of the activity of ADAMTS13 in the sample.