Cadmium chelate complex monoclonal antibody and preparation method and usage thereof

A monoclonal antibody, chelate technology, applied in the direction of anti-animal/human immunoglobulin, measuring devices, instruments, etc., can solve the analysis work can only be carried out in indoor laboratories, labor-intensive, long analysis time, etc. problem, to achieve the effect of fast detection speed, low detection cost and accurate detection results

Inactive Publication Date: 2008-03-12
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, Cd in the environment and agricultural and livestock products 2+ The detection of residues is mainly based on graphite furnace atomic absorption spectrophotometry. This method has the advantages of sensitivity, accuracy, and good selectivity. Large-scale instruments and equipment, the analysis work can only be carried out in the indoor laboratory, which is time-consuming and laborious, and it is difficult to meet the needs of the current detection work

Method used

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  • Cadmium chelate complex monoclonal antibody and preparation method and usage thereof
  • Cadmium chelate complex monoclonal antibody and preparation method and usage thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] (1) Dissolve BSA and KLH in PBS at pH 9.5, respectively, at a concentration of 4 mg / mL;

[0041] (2) 10 mg of chelating agent ITCBE was dissolved in 0.1 mol / L, sodium phosphate buffer solution of pH 9.5, and the concentration was 18 mg / mL;

[0042] (3) Synthesis of the hapten Cd-ITCBE: 1000 μL of 1 mg / mL Cd 2+ Add the standard solution to 2000 μL of 4.6mM chelating agent solution, adjust the pH to 7.0 with ammonia water, and stir for 12 hours at 20°C;

[0043] (4) Synthesis of the immunogen Cd-ITCBE-KLH: take 2 mg of hapten, add 2 mg of KLH, adjust the pH to 8.5 with ammonia water, and react at 20° C. for 12 hours.

[0044] (5) Synthesis of the coated antigen Cd-ITCBE-BSA: take 2 mg of hapten, add 2 mg of BSA, adjust the pH to 8.5 with ammonia water, and react with stirring at 20° C. for 12 h.

[0045] (6) Purification of immunogens and antigens: after the coupling reaction, unreacted small molecular substances were removed by centrifugation using centeicon-30 ultrafi...

Embodiment 2

[0053] (1) Dissolve BSA and KLH in PBS at pH 9.5, respectively, at a concentration of 5 mg / mL.

[0054] (2) Dissolve 10 mg of chelating agent ITCBE in 0.1 mol / L, pH9.5 sodium phosphate buffer solution, the concentration is 20 mg / mL.

[0055] (3) Synthesis of the hapten Cd-ITCBE: 100 μL of 1 mg / mL Cd 2+ The standard solution was added to 200 μL of 4.6 mM chelating agent solution, the pH was adjusted to 7.5 with ammonia water, and the reaction was stirred at 25° C. for 24 h.

[0056] (4) Synthesis of immunogen Cd-ITCBE-KLH: Take 3 mg of hapten, add 3 mg of KLH, adjust the pH to 9.0 with ammonia water, and react with stirring at 25°C for 24 hours.

[0057] (5) Synthesis of the coated antigen Cd-ITCBE-BSA: take 3 mg of hapten, add 3 mg of BSA, adjust the pH to 9.0 with ammonia water, and react with stirring at 25°C for 24 hours.

[0058] (6) Purification of immunogens and antigens: after the coupling reaction, unreacted small molecular substances were removed by centrifugation u...

Embodiment 3

[0070] (1) Dissolve BSA and KLH in PBS at pH 9.5, respectively, at a concentration of 6 mg / mL.

[0071] (2) Dissolve 10 mg of chelating agent ITCBE in 0.1 mol / L sodium phosphate buffer solution with pH 9.5 to a concentration of 22 mg / mL.

[0072] (3) Synthesis of the hapten Cd-ITCBE: 1000 μL of 1 mg / mL Cd 2+ The standard solution was added to 2000 μL of 4.6 mM chelating agent solution, the pH was adjusted to 8.0 with ammonia water, and the reaction was stirred at 30° C. for 36 h.

[0073] (4) Synthesis of immunogen Cd-ITCBE-KLH: Take 4 mg of hapten, add 4 mg of KLH, adjust the pH to 9.5 with ammonia water, and react with stirring at 30° C. for 36 h.

[0074] (5) Synthesis of the coated antigen Cd-ITCBE-BSA: take 4 mg of hapten, add 4 mg of BSA, adjust the pH to 9.5 with ammonia water, and react with stirring at 30°C for 36 hours.

[0075] (6) Purification of immunogens and antigens: after the coupling reaction, unreacted small molecular substances were removed by centrifugat...

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Abstract

The present invention discloses a cadmium chelate monoclonal antibody and a preparation method as well as application of the monoclonal antibody. The monoclonal antibody is of specificity to cadmium -1 - (4 - isothiocyanate-benzyl) - EDTA - bovine serum albumin and the cadmium metal chelate. The preparation method of the cadmium chelate monoclonal antibody is as following: first of all, hapten is prepared and cadmium chelate complete antigen is formed; spleen cells of mouse which are Cd-ITCBE-KLH-immune are combined with SP2 / 0, and the positive clones are screened through a two-step method; after a plurality of times of subcloning, monoclonal antibody that can stably excrete anti-cadmium chelate is obtained. The cadmium chelate monoclonal antibody is used to measure the concentration of cadmium ion through the linear equation: Y = 0.0081X +0.1115, R 2 = 0.9922. The measurement is fast and the cost is low; the measurement result is accurate and is of high sensitivity and strong selectivity; the equipment is simple and can be easily carried for on-spot testing.

Description

technical field [0001] The invention relates to a cadmium chelate monoclonal antibody and a preparation method thereof. Cadmium chelate monoclonal antibody can be applied to Cd in the environment and agricultural and livestock products 2+ residual detection. Background technique [0002] Cadmium (Cadmium, Cd) is one of the most harmful elements that threaten human health. It is a slightly toxic heavy metal, and is listed as the third food pollutant that needs priority research by the Food and Agriculture Organization of the United Nations and the World Health Organization. Cadmium mainly harms the kidneys and liver and can cause pulmonary edema, high blood pressure, anemia, osteomalacia and hyposmia loss. Eating cadmium-containing compounds by mistake can cause acute poisoning, nausea, vomiting, diarrhea, general fatigue, muscle pain, and collapse; long-term consumption of foods containing cadmium can cause body cancer and neonatal deformities. [0003] At present, Cd in ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/18G01N33/577
Inventor 王菊芳李志勇刘功良王小宁
Owner SOUTH CHINA UNIV OF TECH
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