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.

Description

TECHNICAL FIELD[0001]The present invention relates to a kit for preparing a sample for detection of a monoclonal antibody through mass spectrometry; more particularly, to such a kit that enables selective protease reaction to obtain peptide fragments containing the specific sequence of a monoclonal antibody so that even more efficient analysis is achieved.BACKGROUND ART[0002]The hardest challenge in drug discovery is to develop drugs capable of showing high efficacy with minor side effects. For that matter, the current focus is on pharmacokinetics, especially therapeutic drug monitoring (TDM). Whether the prescribed dosage is proper and whether the drug has reached the lesion are indicators for drug seeds to be screened so as to determine which drug seeds need to be dropped at an early stage of development. The TDM information is useful in early-stage drug discovery and clinical testing. Personalized medicine, also referred to as order-made medicine, was anticipated at one time; how...

AI Technical Summary

Benefits of technology

[0034]The kit related to the present invention is used for a process to be conducted prior to liquid chromatography-mass spectrometry (especially a triple quadrupole type). Examples of a mass spectrometer applicable for practicing the present invention are LCMS-8030, LCMS-8040, LCMS-8050 and LCMS-8080 (all made by Shimadzu Corporation). In addition, to analyze the sample pretreated using the kit related to the present invention, LCMS-IT-TOF and LCMS-Q-TOF (both made by Shimadzu Corporation) may be used.

Problems solved by technology

The hardest challenge in drug discovery is to develop drugs capable of showing high efficacy with minor side effects.

Personalized medicine, also referred to as order-made medicine, was anticipated at one time; however, from the viewpoint of medical economics, providing personalized medical technique and medications is virtually impossible except for some wealthy patients.

Antibodies are said to naturally show significantly high molecular specificity and to accumulate in targeted lesions; however, no technology exists to prove such characteristics.

However, there are problems yet to be solved in applications of ELISA: for example, abnormal values may appear since the analyte is not directly measured; it may be time-consuming and costly since antibodies need to be labeled for each target; simultaneous detection of multiple analytes is not available; and so on.

Especially, since cross-reactions with endogenous antibodies may occur when antibody drugs are used, it is hard to obtain accurate detection results.

Moreover, when a neutralizing antibody and the antigen are bound, antigen recognition sites may be blocked to inhibit the use of ELISA.

Moreover, due to interspecies specificity issues, analysis conditions employed during the animal-testing phase by ELISA are often inadequate to use for testing on large animals and human beings.

Since detection-inhibiting matrix components are different in ELISA for determining the drug concentration in lesional tissues, it is necessary to label multiple antibodies to perform pharmacokinetics analysis by ELISA, thereby entailing significant risks such as remarkably high cost, dropout at a later stage of development, and the like.

In the field of medicine, errors of a doctor in slicing a lesion and preserving the sliced pathological section, differences in available facilities, and other issues have led to differences in immunohistochemical staining, thereby oftentimes making it difficult to determine whether the result is positive, false-positive or negative.

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