Preparation of multi-organophosphor universal antibody and universal envelope antigen

Abstract

The invention discloses preparation of various common antibodies of an enzyme-linked immunoadsorption determination reagent kit suitable for analyzing different residues at the same time and common envelope antigen, which comprise preparation methods of common organophosphorus pesticide semiartificial antigen structure, different organophosphorus pesticide common antibodies and common envelope antigen. The organophosphorus pesticide common antibodies and the common envelope antigen prepared by the preparation methods can be used to prepare the enzyme-linked immunoadsorption determination reagent kit suitable for analyzing different organophosphorus pesticide residues at the same time; moreover, the reagent kit can be used for quick detection of different organophosphorus pesticides such as methyl parathion, fenitrothion, fenthion, acephate, Cygon and malathion in water, soil, foodstuff such as vegetables and poisoned samples; in addition, the invention has the advantages of simple preliminary treatment process of samples, simultaneous detection of samples in batches and lower sample detection cost than the prior detection method.

Description

(1) Technical field

[0001] The invention belongs to the field of residual pesticide detection and relates to the preparation of a variety of organophosphate general antibodies and general coating antigens which are suitable for an enzyme-linked immunosorbent assay kit for simultaneous analysis of multiple organophosphate residues. (2) Background technology

[0002] The traditional residue analysis methods of pesticides and their metabolites mainly rely on physical and chemical analysis methods such as gas chromatography (GC), high performance liquid chromatography (HPLC), and mass spectrometry (MS). The chronic and long-term effects of human health and human health are increasingly concerned and worried, and the restrictions on pesticide residues are becoming more and more stringent. New requirements and higher requirements are put forward for the analysis and measurement objects, types, quantities, ranges, indicators, etc. However, the traditional physical and chemical anal...

Examples

Embodiment Construction

[0034] The present invention will be further described below with embodiment:

[0035] Artificial antigen preparation: d aryl phosphorothioate artificial hapten, direct phosphorothioate artificial hapten 60mg, 72mg were dissolved in 5.0ml DMF. Dissolve 0.2 mL of tri-n-butylamine in 5.0 mL of DMF, and add e to d to obtain f. Dissolve 0.16 mL of isobutyl chloroformate in 10 mL of DMF, add dropwise to f at 4°C, and react for 1 hour to obtain g. 120 mg of bovine serum albumin was dissolved in 60 mL of borate buffer solution with pH=8.7 and 0.05 M, then g was added dropwise to it below 18°C, and reacted at room temperature (~20°C) for 12 hours. The obtained reaction solution was put into a dialysis bag, and dialyzed in PH=7.4, 0.01M buffer solution at 4°C for 60 hours, and the PBS dialysate was changed every 8 hours, 1 L each time. Until no hapten UV absorbance (OD 282 ). Accurately measure the volume of the protein solution after dialysis, determine the protein concentration a...

(1) Technical field

[0001] The invention belongs to the field of residual pesticide detection and relates to the preparation of a variety of organophosphate general antibodies and general coating antigens which are suitable for an enzyme-linked immunosorbent assay kit for simultaneous analysis of multiple organophosphate residues. (2) Background technology

[0002] The traditional residue analysis methods of pesticides and their metabolites mainly rely on physical and chemical analysis methods such as gas chromatography (GC), high performance liquid chromatography (HPLC), and mass spectrometry (MS). The chronic and long-term effects of human health and human health are increasingly concerned and worried, and the restrictions on pesticide residues are becoming more and more stringent. New requirements and higher requirements are put forward for the analysis and measurement objects, types, quantities, ranges, indicators, etc. However, the traditional physical and chemical anal...

Examples

Embodiment Construction

[0034] The present invention will be further described below with embodiment:

[0035] Artificial antigen preparation: d aryl phosphorothioate artificial hapten, direct phosphorothioate artificial hapten 60mg, 72mg were dissolved in 5.0ml DMF. Dissolve 0.2 mL of tri-n-butylamine in 5.0 mL of DMF, and add e to d to obtain f. Dissolve 0.16 mL of isobutyl chloroformate in 10 mL of DMF, add dropwise to f at 4°C, and react for 1 hour to obtain g. 120 mg of bovine serum albumin was dissolved in 60 mL of borate buffer solution with pH=8.7 and 0.05 M, then g was added dropwise to it below 18°C, and reacted at room temperature (~20°C) for 12 hours. The obtained reaction solution was put into a dialysis bag, and dialyzed in PH=7.4, 0.01M buffer solution at 4°C for 60 hours, and the PBS dialysate was changed every 8 hours, 1 L each time. Until no hapten UV absorbance (OD 282 ). Accurately measure the volume of the protein solution after dialysis, determine the protein concentration a...

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