Enzyme-linked immunosorbent assay method for detecting cadmium-metallothionein in flesh of shell

An enzyme-linked immunosorbent adsorption and metallothionein technology is applied in the direction of measuring devices, instruments, scientific instruments, etc., to achieve the effect of scientific and reasonable principles, mature technical steps and high sensitivity

Inactive Publication Date: 2010-12-01
GUANGDONG OCEAN UNIVERSITY
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Problems solved by technology

[0006] At home and abroad, there are studies on the extraction and preparation of metallothionein and the enzyme-linked immunosorbent assay. Rabbits are induced to synthesize cadmium-metallothionein antibodies, and then the antibodies are purified to establish enzyme-linked immunosorbent assays for the detection of cadmium metallothionein in shellfish. The research on the indirect detection of cadmium content in shellfish meat is still blank

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Embodiment Construction

[0023] The above-mentioned Sephadex G-50 chromatography is eluted with pH8.60.01mol / L Tris-HCl buffer at an elution rate of 1ml / min, and DEAE-52 column chromatography is buffered with pH8.60.01mol / L Tris-HCl The solution was eluted with a gradient of 0-0.5 mol / L NaCl at a flow rate of 1 ml / min. In the process of antibody induction, 100-200 μg of cadmium-metallothionein was injected at multiple points each time, for a total of six injections. The first three times were emulsified with one volume of Freund's complete adjuvant, and the last three times were emulsified with one volume of Freund's inert adjuvant. Complete adjuvant emulsification. The crudely extracted cadmium-metallothionein antibody was purified by elution of DEAE-Sephadex A50 column with pH7.40.01mol / L PBS solution.

[0024] During the establishment of ELISA, add 80-120 μl of coating solution for the sample to be tested, coat overnight at 2-5°C, wash with 250 μl washing solution for 3-5 times; add 100-300 μl of ...

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Abstract

The invention discloses an enzyme-linked immunosorbent assay method for detecting cadmium-metallothionein in the flesh of a shell. The method comprises the following steps of: inducing the shell to synthesize the cadmium-metallothionein by using cadmium chloride, inducing a rabbit to synthesize an antibody by using purified metallothionein, separating and purifying the antibody, using an electrophoretically pure rabbit anti-shell metallothionein antibody as a primary antibody and a goat anti-rabbit enzyme labelled antibody as a secondary antibody, and assaying the cadmium-metallothionein content of the shell by using the indirect competitive enzyme-linked immunosorbent assay method. The method has the advantages of scientific and rational principle and relatively mature technical process and is used for assaying the cadmium-metallothionein content of the flesh of the shell so as to indirectly reflect the cadmium pollution of the flesh of the shell and provide a fast and sensitive detection method for assaying the heavy metal cadmium in the flesh of the shell.

Description

technical field [0001] The invention belongs to an enzyme-linked immunosorbent assay method for detecting metallothionein in shellfish. Background technique [0002] Due to the increasing mining, smelting, processing and commercial manufacturing activities of heavy metals, many heavy metals such as lead, mercury, cadmium and cobalt have entered the atmosphere, water and soil, causing serious environmental pollution. Among the industrial "three wastes" pollution, especially heavy metals such as cadmium, mercury, lead and other environmental pollution are the most prominent. Aquatic organisms have a strong ability to enrich cadmium, and cadmium in water can reach a very high concentration in the body after being enriched through the food chain. For example, fish can enrich 10 3 ~10 5 times, shellfish enrichment 10 5 ~10 6 times. After cadmium is absorbed by the human body, it forms cadmium protein in the body and selectively accumulates in the kidney and liver. The kidney...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543
Inventor 吴晓萍廖爱琳廖艳吴育廉杨捷梁鹏邓楚津
Owner GUANGDONG OCEAN UNIVERSITY
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