Indirectly racing ELISA detecting method for cadmium ion

A detection method, the technology of cadmium ions, applied in the biological field, can solve the problems of insufficient reporting and the inability to repeatedly synthesize artificial antigens, and achieve the effect of guaranteeing interests, low cost, and sensitive detection

Inactive Publication Date: 2008-06-11
JINAN UNIVERSITY
View PDF0 Cites 15 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There is no report on the immunological detection of heavy metals in China, and the reports in foreign literature are insufficient. Artificial antigens cannot be repeatedly

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
  • Indirectly racing ELISA detecting method for cadmium ion
  • Indirectly racing ELISA detecting method for cadmium ion
  • Indirectly racing ELISA detecting method for cadmium ion

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0034] Example 1

[0035] The indirect competitive ELISA method for detecting cadmium ions by using anti-cadmium ion monoclonal antibody includes the following steps:

[0036] (A) Cadmium ion coupling to hemocyanin (KLH) with iEDTA (i.e. 1-(4-isothiocyanatobenzyl)ethylenediamine-N,N,N',N'-tetraacetic acid), BALB / c mice were immunized with incomplete Freund's adjuvant, spleen cells of the immunized mice were fused with SP2 / 0 myeloma cells, and positive cell lines that stably secreted anti-cadmium ion monoclonal antibody (MAb-Cd) were screened and expanded. Culture, inject cells into mice to induce ascites, and purify to obtain anti-cadmium ion monoclonal antibody.

[0037] (B) The detection antigen Cd-iEDTA-BSA was obtained by coupling cadmium ions to the carrier protein BSA with iEDTA.

[0038] (C) Preparation of antigen pre-coated strips:

[0039] (1) Coating: Dilute Cd-iEDTA-BSA to 0.2 μg / mL with coating buffer (0.05M sodium carbonate buffer, pH 9.6), and add 100 μL per m...

Example Embodiment

[0060] Example 2

[0061] The indirect competitive ELISA method for detecting cadmium ions by using anti-cadmium ion monoclonal antibody includes the following steps:

[0062] (A) Using iEDTA to couple cadmium ions to limpet hemocyanin (KLH), BALB / c mice were immunized with incomplete Freund's adjuvant, and spleen cells of the immunized mice were fused with SP2 / 0 myeloma cells. The positive cell line that stably secretes anti-cadmium ion monoclonal antibody (MAb-Cd) is expanded and cultured, and the cells are injected into mice to induce ascites, and the anti-cadmium ion monoclonal antibody is purified and obtained.

[0063] (B) The detection antigen Cd-iEDTA-BSA was obtained by coupling cadmium ions to the carrier protein BSA with iEDTA.

[0064] (C) Preparation of antigen pre-coated strips:

[0065] (1) Coating: Cd-iEDTA-BSA was diluted to 0.2 μg / mL with coating buffer (0.05M sodium carbonate buffer, pH 9.6), and 100 μL per microwell was added to the micron of polystyrene ...

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 discloses an ELISA method by indirect competition for testing the cadmium ion, which is to add the extracting solution of the seafood samples and the standard fluid of cadmium ion in serial consistency into the millipore of the antigen pre-peridium strips, and then add the monoclonal antibody with the function of anti-cadmium ion into each millipore, blend for incubation, add the antibody with the function of anti-mouse IgG marked by HRP and then make incubation; stop the reaction after the substrate chromogenic cadmium ion, test the absorbency of each millipore by enzyme mark instrument, and calculate the contact in the samples in accordance with the curvilinear regression equation gained from the standard control. By utilizing the gained secretory positive cells of the monoclonal antibody with the function of anti-cadmium ion, the method provided in the invention can prepare in mass production the monoclonal antibody with the function of anti-cadmium ion. The method costs low, which can fast, simply and sensitively test the content of the cadmium ion in the samples. The invention can make fast and sensitive test on the heavy metal cadmium left in the seafood like fishes and shellfishes on a large scale.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a rapid diagnosis method for heavy metal residues in aquatic meat products, in particular to an indirect competitive ELISA method for detecting cadmium ions using monoclonal antibodies against heavy metal cadmium ions. Background technique [0002] Cadmium (Cdmium) atomic number 48 is one of the heavy metals that seriously endanger human health, mainly from cadmium mines and cadmium smelters. Because cadmium and zinc are in the same family and often coexist with zinc, there must be ZnO and CdO in the emissions of zinc smelting. They are highly volatile and can spread thousands of meters away from the pollution source. With the rapid development of industry and agriculture in our country, the pollution of heavy metals in the environment is becoming more and more serious. The poison caused by heavy metal pollution persists and will enter the food chain with the recirculation of soil and...

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): G01N33/577G01N33/543
Inventor 向军俭王建华唐勇黄峙邓宁王宏杨红宇
Owner JINAN UNIVERSITY
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