Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Pharmacokinetic mass spectrometry method of monoclonal antibody medicine

A monoclonal antibody, pharmacokinetic technology, applied in the field of proteomics, can solve the problems of inability to distinguish endogenous protein interference, long development cycle, narrow quantitative range, etc., to achieve simple and easy sample processing and quantitative methods. , Conducive to the promotion of the method, the effect of the simple preparation method

Inactive Publication Date: 2016-05-11
JILIN UNIV
View PDF3 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in monoclonal antibody pharmacokinetic experiments, the ELISA method has a narrow quantitative range (usually one to two orders of magnitude), cannot simultaneously analyze different forms of monoclonal antibody (ie, monoclonal antibody) drugs, and there is a development cycle Long time (half a year to several years), high cost (hundreds of thousands to millions per target drug), inability to distinguish interference from endogenous proteins, etc., greatly affecting the reliability and practicability of ELISA quantification
For the RIA method, the target antibody of some radiolabeled molecules will show different absorption, distribution and metabolism characteristics from the prototype drug; in addition, the radiolabeled monoclonal antibody usually only exists stably for 2 to 3 weeks, and cannot adapt to medium and long-term Monoclonal Antibody Pharmacokinetic Study

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Pharmacokinetic mass spectrometry method of monoclonal antibody medicine
  • Pharmacokinetic mass spectrometry method of monoclonal antibody medicine
  • Pharmacokinetic mass spectrometry method of monoclonal antibody medicine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1: Analysis of monoclonal antibody drugs in rat plasma samples

[0041] Part I: Intravenous Administration in Rats

[0042] A. Rat intravenous administration process

[0043] (1) choose a male rat with a body weight of 200g ± 10g, and eat and drink freely;

[0044] (2) Purchase commercially available monoclonal antibody drug Bevacizumab injection with a concentration of 25 mg / mL;

[0045] (3) According to the conversion of the clinical dosage, administer the monoclonal antibody drug to rats by intravenous injection at a dosage of 50 mg / kg;

[0046] B. Preparation of Rat Plasma Samples

[0047] Different time points (0, 30min, 1h, 2h, 4h, 8h, 12h, 24h, 32h) were selected, and rat whole blood was collected from the vein, and the serum was centrifuged at 13000g for 5min and stored in a -80°C refrigerator.

[0048] Part II: Determination of the monoclonal antibody drug Bevacizumab in rat plasma samples

[0049] A. Plasma sample pretreatment

[0050] (1) Reduct...

Embodiment 2

[0089] Example 2 Rat plasma standard curve sample preparation

[0090] First prepare the monoclonal antibody standard series solution: dilute the monoclonal antibody standard solution stock solution (25 μg / μL) to 5, 10, 40, 100, 500, 1000, 1200 ng / μL with deionized water, respectively.

[0091] Then prepare the rat plasma standard curve sample: proceed in the following 7 steps,

[0092] (1) Reduction of rat plasma standard curve samples,

[0093] Add 2 μL monoclonal antibody standard series solution into the polyethylene tube respectively,

[0094] Add 2 μL rat blank plasma, vortex to mix,

[0095] Add 94 μL of pH buffer containing denaturant, vortex to mix,

[0096] Add 2 μL reducing agent, vortex to mix,

[0097] Incubate at 60°C for 1 hour to obtain a reaction solution;

[0098] (2) Blocking of cysteine,

[0099] Add 0.75 mg of solid cysteine ​​blocking agent to the reaction solution, and vortex to mix;

[0100] Incubate at 25°C in the dark for 40 minutes;

[0101] ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention provides a pharmacokinetic mass spectrometry method of a monoclonal antibody medicine and belongs to the technical field of proteomics. With the adoption of the technical scheme, the pharmacokinetic mass spectrometry method comprises the following steps: selecting a specific peptide fragment obtained by hydrolyzing Bevacizumab with a pancreatic enzyme, wherein the specificity means that the peptide fragment is generated by only hydrolyzing the Bevacizumab with the pancreatic enzyme, and the peptide fragment cannot be generated by hydrolyzing other proteins and protein type medicines through the pancreatic enzyme; based on a liquid chromatograph-tandem mass spectrometry technology, scanning and analyzing the specific peptide fragment by using a parallel reaction monitoring mode, and establishing a stable and reliable LC / MS / MS quantifying method of the specific peptide fragment. The pharmacokinetic mass spectrometry method of the monoclonal antibody medicine has the characteristics of high flux, high sensitivity, good repeatability and the like; used reagents and medicines are low in price and easy to obtain and a preparation method is simple, so that the popularization of the method is facilitated; a sample treatment process and a quantifying method are simple and feasible, and the method has guide and reference meanings on qualitative and quantitative researches on the other protein type medicines and polypeptide medicines.

Description

technical field [0001] The invention belongs to the technical field of proteomics, and relates to a pharmacokinetic mass spectrometry analysis method for a therapeutic monoclonal antibody drug Bevacizumab. Background technique [0002] Monoclonal antibody drugs are a class of protein drugs with special therapeutic effects based on the structure of γ-type immunoglobulin (ImmunoglobulinG), and have been successfully used in the clinical treatment of various diseases such as cancer, autoimmune diseases, viral infections, and central nervous disorders. treat. Compared with traditional small molecule drugs or traditional Chinese medicine preparations, monoclonal antibody drugs have clear pharmacological activity and excellent pharmacokinetic properties; Elimination half-lives range from weeks to months on average. Therefore, it is of great significance to establish standardized and reliable analytical methods in time and to conduct pharmacokinetic studies of monoclonal antibody...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): G01N30/02G01N30/06
CPCG01N30/02G01N30/06G01N33/6848G01N2030/067
Inventor 胡良海丛宇婷杨波张章顾景凯
Owner JILIN UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com