	US 12,435,376 B2
	Method for high-sensitivity and high-specificity detection of biomolecules by using mass spectrometry
Tae Geol Lee, Daejeon (KR); and Hee-Kyung Na, Daejeon (KR)
Assigned to Korea Advanced Institute of Science and Technology, Daejeon (KR)
Appl. No. 17/271,699
Filed by Korea Research Institute of Standards and Science, Daejeon (KR)
PCT Filed Sep. 3, 2018, PCT No. PCT/KR2018/010217 § 371(c)(1), (2) Date Feb. 26, 2021, PCT Pub. No. WO2020/045713, PCT Pub. Date Mar. 5, 2020.
Claims priority of application No. 10-2018-0103995 (KR), filed on Aug. 31, 2018.
Prior Publication US 2021/0318304 A1, Oct. 14, 2021
Int. Cl. C12Q 1/6886 (2018.01); G01N 33/53 (2006.01)

CPC C12Q 1/6886 (2013.01) [G01N 33/5308 (2013.01); G01N 2610/00 (2013.01); G01N 2800/00 (2013.01)]

11 Claims

1. A method for quantification of biomolecules by using mass spectrometry, the method comprising:

generating a precipitate to quantify the biomolecules by:

i) fixing a first probe molecule in a metal;

ii) reacting the biomolecules with the fixed first probe molecule to form a biomolecule complex;

iii) reacting a second probe molecule with the biomolecule complex, which is formed by reacting the biomolecules with the fixed first probe molecule, so that the second probe molecule binds to the biomolecule complex; and

iv) binding an enzyme conjugate to the bound second probe molecule, wherein the enzyme conjugate is a conjugate in which a material interacting with the second probe molecule is conjugated with the enzyme;

reacting a substrate with the enzyme of the enzyme conjugate to form an insoluble product; and

quantifying the insoluble product by using mass spectrometry,

wherein the biomolecules are microRNA (miRNA).