US 12,305,217 B2
Ex vivo protease activity detection for disease detection/diagnostic, staging, monitoring and treatment
Faycal Touti, Belmont, MA (US); Wendy Winckler Adamovich, Melrose, MA (US); Sophie Cazanave, Cambridge, MA (US); Mehar Cheema, Medford, MA (US); and Robert S. Langer, Newton, MA (US)
Assigned to Sunbird Bio, Inc., Cambridge, MA (US)
Filed by Sunbird Bio, Inc., Cambridge, MA (US)
Filed on Dec. 9, 2022, as Appl. No. 18/063,812.
Application 18/063,812 is a continuation of application No. 17/573,129, filed on Jan. 11, 2022, granted, now 11,604,193.
Application 17/573,129 is a continuation in part of application No. PCT/US2021/049948, filed on Sep. 10, 2021.
Claims priority of provisional application 63/077,525, filed on Sep. 11, 2020.
Prior Publication US 2023/0175037 A1, Jun. 8, 2023
Int. Cl. C12Q 1/37 (2006.01); G01N 21/64 (2006.01); G01N 33/542 (2006.01); G01N 33/573 (2006.01); G01N 33/58 (2006.01)
CPC C12Q 1/37 (2013.01) [G01N 21/6428 (2013.01); G01N 33/542 (2013.01); G01N 33/573 (2013.01); G01N 33/582 (2013.01); G01N 2021/6432 (2013.01); G01N 2021/6439 (2013.01); G01N 2021/6441 (2013.01); G01N 2333/95 (2013.01); G01N 2800/085 (2013.01); G01N 2800/26 (2013.01); G01N 2800/7028 (2013.01)] 26 Claims
OG exemplary drawing
 
1. A method, comprising:
a. collecting a body fluid sample from a subject;
b. contacting the body fluid sample from the subject with one or more synthetic molecules ex vivo,
i. wherein said one or more synthetic molecules comprise a plurality of reporters and a plurality of cleavable peptide linkers, wherein a first cleavable peptide linker of said plurality of cleavable peptide linkers is coupled to a first reporter of said plurality of reporters, wherein a second cleavable peptide linker of said plurality of cleavable peptide linkers is coupled to a second reporter of said plurality of reporters, and wherein the reporter of said plurality of reporters is directly connected to the cleavable peptide linker of said plurality of cleavable peptide linkers through a covalent bond,
ii. wherein said one or more synthetic molecules react with a plurality of extracellular or circulating enzymes from said body fluid sample, wherein a first enzyme of said plurality of extracellular or circulating enzymes cleaves said first cleavable peptide linker and releases said first reporter from said one or more synthetic molecules, wherein said release of said first reporter forms a first detectable signal, and wherein a second enzyme of said plurality of extracellular or circulating enzymes cleaves said second cleavable peptide linker and releases said second reporter from said one or more synthetic molecules, wherein said release of said second reporter forms a second detectable signal; and
c. detecting said first detectable signal or said second detectable signal.