Kit for preparing sample for detecting monoclonal antibody

Abstract

A sample preparation kit related to the present invention provides a significantly versatile analytical technique that is not affected by the diversity of antibodies, difference in species, matrix and the like. For preparing a sample to be used for detection of a monoclonal antibody through high-performance liquid chromatography-mass spectrometry (LC-MS), the kit includes a porous body for immobilizing a monoclonal antibody to be detected; nanoparticles with an immobilized protease; a reaction vessel for selectively digesting the monoclonal antibody by bringing the porous body and nanoparticles into contact; a buffer to be introduced into the reaction vessel along with the nanoparticles and porous body so that a protease reaction is carried out; and a filtration membrane to remove the porous body and nanoparticles after the proteolysis so as to extract the reaction product and the buffer.

1 - 11 . (canceled) 
     
     
         12 : A kit for preparing a sample for detection of a monoclonal antibody through high-performance liquid chromatography-mass spectrometry, comprising:
 a porous body that immobilizes a monoclonal antibody;   nanoparticles having an immobilized protease;   a reaction vessel that selectively proteolyses the monoclonal antibody by bringing the porous body and nanoparticles into contact;   a buffer to be introduced into the reaction vessel along with the nanoparticles and porous body such that a protease Proteolysis reaction is carried out; and   a filtration membrane that removes the porous body and nanoparticles after the Proteolysis reaction and extracts a Proteolysis reaction product and the buffer,   wherein the filtration membrane functions as a bottom of the reaction vessel which hardly permeates the buffer and a peptide produced through the protease Proteolysis reaction under a condition that no pressure or centrifugal force is applied and permeates the buffer and peptide under a condition that pressure or centrifugal force is applied.   
     
     
         13 : The kit according to  claim 12 , wherein the filtration membrane is made of polyvinylidene difluoride. 
     
     
         14 : A kit for processing multiple samples simultaneously, comprising
 the kit of  claim 12 ,   wherein housing material of the filtration membrane is made of a polyacrylonitrile resin.   
     
     
         15 : A kit for detecting a monoclonal antibody through high-performance liquid chromatography-mass spectrometry, comprising:
 the kit of  claim 12 ; and   instructions for mass spectrometry conditions to detect a monoclonal antibody.   
     
     
         16 : The kit according to  claim 12 , further comprising:
 at least one internal standard peptide having an amino-acid sequence specific to a target monoclonal antibody.   
     
     
         17 : The kit according to  claim 16 , wherein the target monoclonal antibody is trastuzumab, trastuzumab-DM1, bevacizumab or rituximab, and the internal standard peptide has at least one amino-acid sequence selected from SEQ ID NO: 1˜47. 
     
     
         18 : A non-transitory computer readable recording medium, wherein the non-transitory computer readable recording medium is used in a method for detecting a monoclonal antibody conducted by bringing a porous body with a target monoclonal antibody immobilized in its pores and nanoparticles with an immobilized protease into contact with each other in a liquid so as to perform selective protease Proteolysis of the monoclonal antibody, and by analyzing the obtained peptide fragments through high-performance liquid chromatography-mass spectrometry, the non-transitory computer readable recording medium having stored thereon data for executing the high-performance liquid chromatography-mass spectrometry, and the data include a parent ion, fragment ion, expected retention time, and voltage at each quadrupole of a triple quadrupole mass spectrometer to be applied on at least one peptide obtained through protease Proteolysis of the monoclonal antibody. 
     
     
         19 : A method package for detecting a monoclonal antibody through high-performance liquid chromatography-mass spectrometry, comprising:
 the recording medium of  claim 18 ; and   instructions for the recording medium.   
     
     
         20 : The recording medium according to  claim 18 , wherein the monoclonal antibody is at least one of trastuzumab, trastuzumab-DM1, bevacizumab and rituximab. 
     
     
         21 : The recording medium according to  claim 18 , wherein the data are related to a peptide having at least one amino-acid sequence selected from SEQ ID NO: 1˜47. 
     
     
         22 : The method package according to  claim 19 , wherein the monoclonal antibody is at least one of trastuzumab, trastuzumab-DM1, bevacizumab and rituximab. 
     
     
         23 : The method package according to  claim 19 , wherein the data are related to a peptide having at least one amino-acid sequence selected from SEQ ID NO: 1˜47. 
     
     
         24 : A kit for processing multiple samples simultaneously, comprising
 the kit of  claim 13 ,   wherein housing material of the filtration membrane is made of a polyacrylonitrile resin.   
     
     
         25 : A kit for detecting a monoclonal antibody through high-performance liquid chromatography-mass spectrometry, comprising:
 the kit of  claim 13 ; and   instructions for mass spectrometry conditions to detect a monoclonal antibody.   
     
     
         26 : The kit according to  claim 13 , further comprising:
 at least one internal standard peptide having an amino-acid sequence specific to a target monoclonal antibody.   
     
     
         27 : The kit according to  claim 26 , wherein the target monoclonal antibody is trastuzumab, trastuzumab-DM1, bevacizumab or rituximab, and the internal standard peptide has at least one amino-acid sequence selected from SEQ ID NO: 1˜47.