Preparation method and application of NDM-1 monoclonal antibody

A monoclonal antibody, NDM-1 technology, applied in the field of molecular biology, can solve the problems of low purity titer, poor specificity, and long time consumption, and achieve the effect of high purity, high sensitivity, and strong specificity

Active Publication Date: 2021-07-23
深圳市龙华区疾病预防控制中心
View PDF3 Cites 6 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] NDM-1 can hydrolyze almost all β-lactam antibiotics, including the most effective carbapenem antibiotics, which brings great challenges to clinical treatment and hospital infection prevention and control
However, the existing NDM-1 detection method has poor specificity and takes a long time, which delays the diagnosis and treatment of patients; the monoclonal antibody of NDM-1 protein in the prior art has low purity and low titer and poor specificity

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
  • Preparation method and application of NDM-1 monoclonal antibody
  • Preparation method and application of NDM-1 monoclonal antibody
  • Preparation method and application of NDM-1 monoclonal antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0042] 1. Main experimental equipment and reagents

[0043] Centrifuge: Type 5702, Type 5418, Eppendorf.

[0044] Micro-dosing gun: Gilson Company.

[0045] Water bath: DKB-8A, Jinghong Company.

[0046] SDS-PAGE electrophoresis system: Mini product, Bio-Rad company.

[0047] Western Blotting transfer system: Mini product, Bio-Rad company.

[0048] Microplate reader: Epoch, BioTek.

[0049] Freund's complete adjuvant and incomplete adjuvant, Ig class and subclass secondary antibodies, dimethyl sulfoxide (DMSO), and PEG SOLUTION were all purchased from Sigma.

[0050] HPR-labeled goat anti-mouse IgG antibody was purchased from Southern Biotech, USA.

[0051] DMEM, HAT, penicillin, and fetal bovine serum were purchased from Gibco.

[0052] PVDF membranes were purchased from Millipore Corporation.

[0053] Pre-stained protein marker was purchased from Thermo Company.

[0054] Bovine serum albumin (BSA) was purchased from Shanghai Sangong Company.

[0055] ECL chromogenic...

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

PropertyMeasurementUnit
molecular weightaaaaaaaaaa
Login to view more

Abstract

The invention discloses a preparation method and application of an NDM-1 monoclonal antibody. The preparation method comprises the following steps: synthesizing a Klebsiella pneumoniae NDM-1 gene, expressing and purifying NDM-1 protein, taking recombinant NDM-1 protein as immunogen, immunizing Balb / c mice, taking spleen cells of the immunized mice to fuse with myeloma NS-1 cells of the mice, screening hybridoma cells secreting specific McAb through an indirect ELISA method, inducing the mice to generate ascites by using a hybridoma cell strain, purifying antibodies by using protein G affinity chromatography to finally obtain seven NDM-1 protein monoclonal antibodies, and sequencing to obtain amino acid sequences of four monoclonal antibodies. A foundation is laid for establishing an NDM-1 detection method.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a preparation method and application of an NDM-1 monoclonal antibody. Background technique [0002] New Delhi metallo-beta-lactamase (NDM-1) is a newly discovered metalloenzyme (MBL) in 2008. It was named after it was first discovered in New Delhi, India. It was isolated from an Indian patient in Sweden. Carrier strains were Klebsiella pneumoniae and Escherichia coli. Subsequently, reports of NDM-1 drug-resistant bacteria appeared in more than 20 countries around the world. More and more data show that NDM-1-producing drug-resistant bacteria have spread globally and may become a threat to human health. The huge hidden dangers caused a global panic. According to reports, NDM-1 widely exists in Gram-negative bacilli, mainly distributed in Enterobacteriaceae and Acinetobacter bacteria, and has been found in Enterobacteriaceae Escherichia, Citrobacter, Salmonella NDM-1 resistant b...

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
Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/12C12N5/20A61K39/42A61P31/04
CPCC07K16/1203A61P31/04A61K2039/505C07K2317/56
Inventor 林启辉汪涛任燕许少坚邬玉兰刘丽
Owner 深圳市龙华区疾病预防控制中心
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap