US 12,216,126 B2
Kit for preparing sample for detecting monoclonal antibody
Takashi Shimada, Kyoto (JP); and Noriko Iwamoto, Kyoto (JP)
Assigned to SHIMADZU CORPORATION, Kyoto (JP)
Filed by SHIMADZU CORPORATION, Kyoto (JP)
Filed on Nov. 16, 2021, as Appl. No. 17/455,089.
Application 17/455,089 is a continuation of application No. 15/556,813, abandoned, previously published as PCT/JP2015/085459, filed on Dec. 18, 2015.
Claims priority of application No. 2015-047729 (JP), filed on Mar. 10, 2015.
Prior Publication US 2022/0137062 A1, May 5, 2022
Int. Cl. G01N 33/68 (2006.01); C12N 11/02 (2006.01); C12Q 1/00 (2006.01); C12Q 1/37 (2006.01); G01N 1/10 (2006.01); G01N 30/06 (2006.01); G01N 30/72 (2006.01); G01N 33/577 (2006.01)
CPC G01N 33/6848 (2013.01) [G01N 1/10 (2013.01); G01N 30/06 (2013.01); G01N 30/7233 (2013.01); G01N 33/577 (2013.01); C12N 11/02 (2013.01); C12Q 1/00 (2013.01); C12Q 1/37 (2013.01); G01N 2030/067 (2013.01); G01N 2333/901 (2013.01)] 20 Claims
 
1. A method for preparing a sample for detection of a monoclonal antibody through high-performance liquid chromatography-mass spectrometry, comprising:
providing porous bodies having pores and a target monoclonal antibody immobilized in the pores of the porous bodies, and nanoparticles having an immobilized protease;
bringing the porous bodies and the nanoparticles into contact with each other in a buffer in a reaction vessel comprising a filtration membrane such that the target monoclonal antibody in the porous bodies and the immobilized protease on the nanoparticles undergo a selective protease proteolysis reaction in the reaction vessel; and
applying pressure or centrifugal force to the reaction vessel containing the buffer such that the buffer and digested peptide fragments produced through the protease proteolysis reaction permeate through the filtration membrane and that the filtration membrane prevents permeation of the porous bodies and the nanoparticles,
wherein the filtration membrane has a pore size set in a range that allows permeation of the digested peptide fragments produced through the protease proteolysis reaction and prevents the permeation of the porous bodies and the nanoparticles.