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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
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  • 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 synthesized based on it. Therefore, it is of great significance to develop an enzyme-linked detection method for heavy metals with independent intellectual property rights.

Method used

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  • 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

Embodiment 1

[0035] Utilize anti-cadmium ion monoclonal antibody to detect the indirect competition ELISA method of cadmium ion, comprises the steps:

[0036] (A) Cadmium ions were coupled to hemocyanin (KLH) using iEDTA (i.e., 1-(4-isothiocyanobenzyl)ethylenediamine-N,N,N',N'-tetraacetic acid), Mix Freund's incomplete adjuvant to immunize BALB / c mice, take the splenocytes of the immunized mice and fuse them with SP2 / 0 myeloma cells, screen out positive cell lines that stably secrete anti-cadmium ion monoclonal antibody (MAb-Cd) and expand Cultured, injected cells into mice to induce ascites, and purified to obtain monoclonal antibodies against cadmium ions.

[0037] (B) Using iEDTA to couple cadmium ions to the carrier protein BSA to obtain the detection antigen Cd-iEDTA-BSA.

[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, pH9.6), add 100 μL per microwell into a polystyrene ...

Embodiment 2

[0061] Utilize anti-cadmium ion monoclonal antibody to detect the indirect competition ELISA method of cadmium ion, comprises the steps:

[0062] (A) Use iEDTA to couple cadmium ions to hemocyanin (KLH), mix Freund's incomplete adjuvant to immunize BALB / c mice, take splenocytes from immunized mice and fuse SP2 / 0 myeloma cells, and screen out Stable positive cell lines secreting anti-cadmium ion monoclonal antibody (MAb-Cd) were expanded and cultured, the cells were injected into mice to induce ascites, and the anti-cadmium ion monoclonal antibody was purified.

[0063] (B) Using iEDTA to couple cadmium ions to the carrier protein BSA to obtain the detection antigen Cd-iEDTA-BSA.

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

[0065] (1) Coating: Dilute Cd-iEDTA-BSA to 0.2 μg / mL with coating buffer (0.05M sodium carbonate buffer, pH9.6), add 100 μL per microwell to the microwell of polystyrene microwell plate Wells were coated overnight at 4°C.

[0066] (2) Washing:...

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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

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Application Information

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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
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