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

Molybdenum ion detection kit based on direct competitive ELISA (enzyme-linked immunosorbent assay) and application of kit

An enzyme-linked immunosorbent assay and detection kit technology, which can be used in measurement devices, instruments, scientific instruments, etc., and can solve problems such as long detection time.

Active Publication Date: 2017-06-13
HENAN INST OF SCI & TECH
View PDF3 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the research on the immunological detection technology of heavy metal molybdenum ion pollution at home and abroad is very active. Among them, the patent with the notification number CN103412127B discloses an enzyme-linked immunosorbent assay kit for detecting hexavalent molybdenum ions and its composition and detection method. The detection sensitivity is 1ng / mL, and the detection time is longer

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
  • Molybdenum ion detection kit based on direct competitive ELISA (enzyme-linked immunosorbent assay) and application of kit
  • Molybdenum ion detection kit based on direct competitive ELISA (enzyme-linked immunosorbent assay) and application of kit
  • Molybdenum ion detection kit based on direct competitive ELISA (enzyme-linked immunosorbent assay) and application of kit

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0043] In the present invention, the preparation method of the detection plate coated with goat anti-mouse IgG secondary antibody preferably comprises the following steps:

[0044] Ⅰ. Add 100-150 μL / well of goat anti-mouse IgG secondary antibody coating solution with a coating concentration of 50-200 μg / mL to the detection wells of the detection plate, and incubate for the first time at 35-40°C for 1.5-3 hours. After incubation, Remove the coating solution, and wash the plate for the first time with the washing solution for 2 to 5 times;

[0045]Ⅱ. Add 200-300 μL / well of pig negative serum with a mass concentration of 3-6% to the detection plate obtained in the above step 1, and incubate for the second time at 35-40°C for 1.5-3 hours, and remove the coating solution after incubation , washing the plate for the second time with washing solution for 2-5 times, and drying the detection plate naturally at 23-27° C. to obtain a detection plate coated with goat anti-mouse IgG second...

Embodiment 1

[0106] Synthesis of Mo-ITCBE-cBSA immunogen ( figure 1 )

[0107] (1) Weigh 10mg ITCBE and dissolve it in 1mL dimethyl sulfoxide (DMSO) to form a metal chelating agent solution; weigh 8.38mg molybdic acid (H 2 MoO 4 ) was dissolved in 1mL of HEPES buffer at pH 8.0 (has no toxic effect on cells. It is a hydrogen ion buffer that can control a constant pH range for a long time) (10mmol / L) to form Mo 6+ solution; the metal chelating agent solution and Mo 6 + The solutions were mixed, and the pH value was adjusted to 7.0 with NaOH, and then reacted on a shaker at room temperature for 12 hours to form the Mo-ITCBE chelate hapten.

[0108] (2) Weigh 66 mg of BSA and 11.6 mg of EDC and dissolve them in 5 mL of PBS buffer, slowly add 7 mg of ethylenediamine (dissolved in 3 mL of PBS + DMF solution in advance) under stirring conditions, and shake at 37 ° C for 2 hours. The reaction solution was dialyzed against PBS for 4 days, and the activated carrier protein BSA was lyophilized a...

Embodiment 2

[0111] Preparation of anti-molybdenum ion monoclonal antibody

[0112] The described anti-molybdenum ion-specific monoclonal antibody is prepared by immunizing Balb / C mice with Mo-ITCBE-cBSA immunogen, and is realized by the following steps:

[0113] (1) Mice immunization: five female Balb / C mice aged 6-8 weeks were immunized with Mo-ITCBE-cBSA at a dose of 60 μg / mouse and a volume of 0.2 mL. The immunogen diluted with PBS was completely emulsified with an equal volume of FCA for the first immunization, and then boosted every 4 weeks, and emulsified with FIA. 7 days after immunization for 5 times, the blood was collected by docking the tail to separate the serum, and the mice with high titer and good blocking effect were screened by indirect ELISA and indirect competitive ELISA (ciELISA) as spare mice for fusion. 3 days before the fusion, the hyperimmunized mice were injected with 50 μg of immunogen in the tail vein and intraperitoneally, each with a volume of 100 μL.

[011...

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 molybdenum ion detection kit based on direct competitive ELISA (enzyme-linked immunosorbent assay). The kit comprises a detection plate coated with a goat anti-mouse IgG secondary antibody, an enzyme-labeled Mo-ITCBE chelate hapten with the mass concentration being 20-50 mu g / mL, an anti-molybdenum-ion monoclonal antibody solution with the mass concentration being 10-20 mu g / mL, 1.0-3.0 mol / L of a stop buffer, a sample diluent, a substrate color developing solution, a scrubbing solution, a molybdenum ion standard solution and an EDTA (ethylene diamine tetraacetic acid) treatment solution with the mass concentration being 10-20 mg / mL, wherein the detection plate is subjected to vacuum sealed packaging; the coating concentration of the goat anti-mouse IgG secondary antibody is 50-200 mu g / mL; the anti-molybdenum-ion monoclonal antibody solution is prepared from a Mo-ITCBE-cBSA immunogen immune Balb / C mouse. The kit has the characteristics of high sensitivity and high specificity and has the advantage of short detection time.

Description

technical field [0001] The invention belongs to the technical field of environmental detection, and in particular relates to a kit for detecting molybdenum ions based on a direct competitive enzyme-linked immunosorbent assay and a preparation method and application thereof. Background technique [0002] As a transition metal element, molybdenum plays an important role in organisms. It has a catalytic effect in the process of nitrogen fixation in organisms. It plays the role of transferring electrons and catalyzing substrates in the redox reaction process in the body. It participates in xanthine oxidase / Composition of dehydrogenase, aldehyde oxidase and sulfite oxidase, also has the function of maintaining global carbon, sulfur and nitrogen metabolism balance. In the natural state, molybdenum is not harmful to organisms, but because molybdenum is widely used in special steel, machinery, petrochemical industry, national defense aviation, electronic appliances and nuclear indu...

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): G01N33/577G01N33/531
CPCG01N33/531G01N33/577
Inventor 姜金庆王磊柳东阳贺永惠张慧辉王自良杨雪峰李广领齐永华张海棠黄华国
Owner HENAN INST OF SCI & TECH
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