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Quantitative Methods for Monoclonal Antibodies

A monoclonal antibody and porous body technology, applied in biochemical equipment and methods, microbe determination/inspection, measuring devices, etc., can solve time-consuming and cost-intensive problems

Active Publication Date: 2020-11-06
SHIMADZU SEISAKUSHO CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Conventionally, ELISA (Enzyme-Linked ImmunoSorbent Assay) is known as the most general technique for quantifying proteins such as antibodies, but there are many problems such as time-consuming and cost

Method used

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  • Quantitative Methods for Monoclonal Antibodies
  • Quantitative Methods for Monoclonal Antibodies
  • Quantitative Methods for Monoclonal Antibodies

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0172] The present invention will be further specifically described based on the following examples, but the present invention is not limited by the examples.

[0173] [Immobilization of protease]

[0174] Protease was immobilized on microparticles by the following steps.

[0175] 1. FG bead washing

[0176] 200 mg of FG beads NHS (manufactured by Tamagawa Seiki Co., Ltd.) was centrifuged (15000 g x 5 minutes, 4° C.), and the supernatant for storage was removed with isopropanol. The supernatant also contains sediment and even floating matter, which should be carefully removed.

[0177] 2. Enzyme Preparation

[0178] 1 mg of Trypsin Gold (manufactured by Promega Corporation) or 5 mg of Trypsin TPCK (manufactured by Sigma-Aldrich Co. LLC.) was opened and dissolved in 25 mM HEPES-NaOH, pH 7.0, cooled to 4°C. After dissolving, transfer to a 50ml centrifuge tube and place on ice. The enzyme was co-washed once with 25mM HEPES-NaOH, pH 7.0, and recovered as much as possible. Th...

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PUM

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Abstract

The present invention provides a method for quantifying a monoclonal antibody, comprising the steps of: contacting a porous body in which a monoclonal antibody to be measured is immobilized in a pore with microparticles immobilized with a specific protease, and quantifying the monoclonal antibody. a step of selective protease digestion; and a step of detecting a peptide fragment obtained by the aforementioned digestion comprising amino acids derived from the CDR2 region of the heavy chain or light chain of the aforementioned monoclonal antibody.

Description

technical field [0001] The present invention relates to a method for quantifying monoclonal antibodies using mass spectrometry, and more specifically, to a method for using a specific enzyme for selective digestion, the method comprising the steps of: selecting a peptide fragment containing a specific sequence of a monoclonal antibody and, a step of detecting the resulting peptide fragments by mass spectrometry. Background technique [0002] In order to develop and administer pharmaceutical agents with few side effects and high efficacy, attention has been paid to pharmacokinetics, especially therapeutic drug monitoring (TDM). Through concentration monitoring, it is possible to confirm whether the administered drug is in an appropriate amount and whether it has reached the lesion site, evaluate the effectiveness of the drug, and adjust the dose appropriately. In addition, regarding the effectiveness of molecularly targeted drugs that have become mainstream in the fields of ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/62C12Q1/37
CPCG01N27/62C12Q1/37C12Y304/21001C12Y304/21004G01N30/7233G01N33/94G01N2333/7158
Inventor 嶋田崇史岩本典子滨田哲畅
Owner SHIMADZU SEISAKUSHO CO LTD
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