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Heavy metal Cu<2+> complete antigen and preparation method thereof

A technology of complete antigens and heavy metals, which is applied in the preparation methods of peptides, chemical instruments and methods, animal/human proteins, etc., to achieve the effects of broad social benefits, rapid detection tools, and good economic value.

Inactive Publication Date: 2010-07-14
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, Cu 2+ complete antigen of Cu 2+ Specific monoclonal antibody and Cu based on it 2+ Immunological detection method has not been reported yet

Method used

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  • Heavy metal Cu&lt;2+&gt; complete antigen and preparation method thereof
  • Heavy metal Cu&lt;2+&gt; complete antigen and preparation method thereof
  • Heavy metal Cu&lt;2+&gt; complete antigen and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Weigh 2mg Cu(NO 3 ) 2 Dissolve in 50μL ultrapure water, 4mg ITCBE in 50μL DMSO, 20mg carrier protein KLH or BSA, OVA in 1mL 100mM HEPES buffer solution (pH9.5). Cu(NO 3 ) 2 The solution was added dropwise to the ITCBE solution, and reacted for 3 hours under constant stirring to form Cu-ITCBE hapten. The Cu-ITCBE solution was added dropwise to 0.5 mL carrier protein solution, and reacted at room temperature for 24 h with shaking. Transfer the antigen to an ultrafiltration tube with a molecular weight cut-off of 30000dal, centrifuge at 7500rpm for 10min, wash 6 times with 100mM HEPES buffer solution (pH7.5), then dilute the antigen with 100mM HEPES buffer solution (pH7.5), and pack to obtain Cu 2+ Complete antigen, stored at -20°C.

Embodiment 2

[0038] Weigh 1mg Cu(NO 3 ) 2 Dissolve in 50μL ultrapure water, 2mg ITCBE dissolve in 50μL DMSO, 10mg carrier protein KLH or BSA, OVA dissolve in 1mL 100mM HEPES buffer solution (pH9.5). Cu(NO 3 ) 2 The solution was added dropwise to the ITCBE solution, and reacted for 3 hours under constant stirring to form Cu-ITCBE hapten. The Cu-ITCBE solution was added dropwise to 0.5 mL carrier protein solution, and reacted at room temperature for 24 h with shaking. Transfer the antigen to an ultrafiltration tube with a molecular weight cut-off of 30000dal, centrifuge at 7500rpm for 10min, wash 6 times with 100mM HEPES buffer solution (pH7.5), then dilute the antigen with 100mM HEPES buffer solution (pH7.5), and pack to obtain Cu 2+ Complete antigen, stored at -20°C.

Embodiment 3

[0040] Weigh 2mg Cu(NO 3 ) 2 Dissolve in 50μL ultrapure water, 4mg ITCBE is dissolved in 50μL DMSO, 20mg carrier protein KLH or BSA, OVA is dissolved in 1mL 50mM carbonate buffer (pH9.5). Cu(NO 3 ) 2 The solution was added dropwise to the ITCBE solution, and reacted for 3 hours under constant stirring to form Cu-ITCBE hapten. The Cu-ITCBE solution was added dropwise to 0.5 mL carrier protein solution, and reacted at room temperature for 24 h with shaking. Transfer the antigen to an ultrafiltration tube with a molecular weight cut-off of 30000dal, centrifuge at 7500rpm for 10min, wash 6 times with 15mM ammonium citrate buffer (pH7.5), then dilute the antigen with 15mM ammonium citrate buffer (pH7.5), Subpackage, get Cu 2+ Complete antigen, stored at -20°C.

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Abstract

The invention discloses a heavy metal Cu<2+> complete antigen and a preparation method thereof. The preparation method comprises the following steps: Cu<2+> is reacted with a bifunctional chelating agent to form a hapten; and a Cu<2+>-chelating agent hapten is reacted with a carrier protein to form a Cu<2+>-chelating agent-carrier protein complete antigen. The antigen has better immunogenicity; and an immune mice with the antigen can be used for preparing a Cu<2+> specific monoclonal antibody and further preparing a rapid Cu<2+> detection ELISA (enzyme-linked immuno sorbent assay) kit of various heavy metals and colloidal gold immunochromatography assay test paper.

Description

Technical field: [0001] The invention discloses a heavy metal Cu 2+ The complete antigen and its preparation method belong to the technical field of immunological method detection. Background technique: [0002] At present, more than 20 kinds of heavy metals are known to be highly toxic, among which lead (Pb), cadmium (Cd), mercury (Hg), copper (Cu), chromium, cobalt, nickel, tin, vanadium, etc. are used in agricultural and livestock products There are varying degrees of residues in the chemical, which pose a serious threat to human health. Therefore, all countries in the world attach great importance to the control and monitoring of heavy metal pollution in the environment and agricultural products. Traditional heavy metal detection methods such as: atomic absorption spectrometry, inductively coupled plasma emission spectrometry, etc. require large and expensive analytical instruments, which cannot be used for on-site detection, and are difficult to adapt to the environme...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/765C07K14/77C07K14/795C07K1/113
Inventor 孔涛刘国文李小兵王哲张焱唐佳佳
Owner JILIN UNIV
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