Simplified monoclonal antibody quantification method

Patent metadata

Field	Value
Publication number	US-11209392-B2
Application number	US-201716954649-A
Country	US
Kind code	B2
Filing date	Dec 28, 2017
Priority date	Dec 28, 2017
Publication date	Dec 28, 2021
Grant date	Dec 28, 2021

Abstract

The present invention provides a method for detecting a monoclonal antibody in a sample, the method comprising: (a) a step of capturing and immobilizing, in pores of a porous body, the monoclonal antibody in the sample; (b) a step of performing selective protease digestion of the monoclonal antibody for 30 min or longer by contacting the porous body having the monoclonal antibody immobilized thereon with nanoparticles having a protease immobilized thereon; and (c) a step of detecting a peptide fragment obtained by the selective protease digestion, using liquid chromatography mass spectrometry (LC-MS), wherein step (b) is carried out under stirring condition for 10 sec to 5 min in the initial reaction stage, and then under static condition. According to the present invention, the detection method of a monoclonal antibody using mass spectrometry is simplified and can be applicable to multisample analysis.

First claim

The invention claimed is:

1. A method for detecting a monoclonal antibody in a sample, the method comprising:
(a) a step of capturing and immobilizing, in pores of a porous body, the monoclonal antibody in the sample;
(b) a step of performing selective protease digestion of the monoclonal antibody for 30 min or longer by contacting the porous body having the monoclonal antibody immobilized thereon with nanoparticles having a protease immobilized thereon; and
(c) a step of detecting a peptide fragment obtained by the selective protease digestion, using liquid chromatography mass spectrometry (LC-MS),
wherein step (b) is carried out under stirring condition for 10 sec to 5 min in a initial reaction stage, and then under static condition.

2. The method according to claim 1 , wherein step (b) further comprises additional stirring for 10 sec to 1 min one or more times in addition to the stirring for 10 sec to 1 min in the initial reaction stage.

3. The method according to claim 1 , wherein the stirring is carried out by pipetting operation performed by an automatic dispenser.

4. The method according to claim 1 , wherein step (b) is carried out in a heating vessel that is set to a predetermined reaction temperature.

5. The method according to claim 1 , wherein the method is capable of quantifying the monoclonal antibody in the sample having an antibody concentration in the range of 0.05 to 300 μg/ml.

Assignees

• Shimadzu Corp

Inventors

• Shimada Takashi
• Iwamoto Noriko
• Takanashi Megumi

Classifications

• H01J49/30 Primary

  using magnetic analysers {, e.g. Dempster spectrometer} · CPC title

• G01N30/72

  Mass spectrometers {(mass spectrometers per se H01J49/00)} · CPC title

• G01N30/06

  Preparation · CPC title

• C12N11/14

  Enzymes or microbial cells immobilised on or in an inorganic carrier · CPC title

• G01N1/28

  Preparing specimens for investigation {including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q}(mounting specimens on microscopic slides G02B21/34; means for supporting the objects or the materials to be analysed in electron microscopes H01J37/20 {; laboratory gas handling apparatus B01L5/00}) · CPC title