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L-lauramide arginine monohydrochloride ethanol ester artificial antigen and specific antibody as well as preparation method and application thereof

A technology of lauramide and arginine, applied in the field of immunochemical analysis, to achieve the effects of high accuracy and sensitivity, avoiding organic solvents and facilitating on-site monitoring

Active Publication Date: 2017-12-01
NINGBO ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But so far, the immunoassay method of LAE has not been reported at home and abroad.

Method used

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  • L-lauramide arginine monohydrochloride ethanol ester artificial antigen and specific antibody as well as preparation method and application thereof
  • L-lauramide arginine monohydrochloride ethanol ester artificial antigen and specific antibody as well as preparation method and application thereof
  • L-lauramide arginine monohydrochloride ethanol ester artificial antigen and specific antibody as well as preparation method and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] 1. Synthesis of artificial antigen

[0030] Based on the fact that lauroyl arginine (LAS) is the main component in the molecular structure of LAE, and can fully expose the structural features and contain carboxyl groups that can be coupled with proteins, it is therefore used as a hapten.

[0031] The structure of the hapten is as follows:

[0032]

[0033] 1.1 Synthesis and purification of immune antigen

[0034] The synthesis of immune antigens adopts the carbodiimide method: dissolve 71.3 mg of lauroyl arginine LAS (about 0.2 mmol) in 1 mL of N,N-dimethylformamide DMF, add 81.6 mg of N-hydroxysuccinate Imide NHS (about 0.6mmol), stirred and reacted at room temperature for 15min, then added 72.9mg of dicyclohexylcarbodiimide DCC (0.3mmol), stirred and reacted overnight at room temperature, centrifuged, took 0.5mL of supernatant and slowly added to In 10 mL of 10 mg / mL bovine serum albumin BSA carbonate buffer solution (CBS, 0.1 mol / L, pH 9.6), the reaction was per...

Embodiment 2

[0046] 2. Synthesis of artificial antigen

[0047] Based on the fact that lauroyl arginine (LAS) is the main component in the molecular structure of LAE, and can fully expose the structural features and contain carboxyl groups that can be coupled with proteins, it is therefore used as a hapten.

[0048] The structure of the hapten is as follows:

[0049]

[0050] 2.1 Synthesis and purification of immune antigen

[0051] The synthesis of immune antigens adopts the carbodiimide method: dissolve 107.0 mg of lauroyl arginine LAS (about 0.3 mmol) in 2 mL of N, N-dimethylformamide DMF, add 122.4 mg of N-hydroxysuccinate Imide NHS (0.9mmol), stirred and reacted at room temperature for 15min, then added 109.4mg of dicyclohexylcarbodiimide DCC (0.45mmol), stirred and reacted overnight at room temperature, centrifuged, took 0.8mL of supernatant and slowly added to 15mL 15mg / mL bovine serum albumin BSA carbonate buffer solution (CBS, 0.1mol / L, pH9.6), magnetic stirring reaction for ...

Embodiment 3

[0057] 3. Synthesis of artificial antigen

[0058] Based on the fact that lauroyl arginine (LAS) is the main component in the molecular structure of LAE, and can fully expose the structural features and contain carboxyl groups that can be coupled with proteins, it is therefore used as a hapten.

[0059] The structure of the hapten is as follows:

[0060] 3.1 Synthesis and purification of immune antigen

[0061] The synthesis of immune antigens adopts the carbodiimide method: 142.6 mg of lauroyl arginine LAS (about 0.4 mmol) is dissolved in 1.5 mL of N, N-dimethylformamide DMF, and 163.2 mg of N-hydroxyl Succinimide NHS (1.2mmol), stirred and reacted at room temperature for 15min, then added 145.8mg of dicyclohexylcarbodiimide DCC (0.6mmol), stirred and reacted overnight at room temperature, centrifuged, took 1mL of supernatant and slowly added to 20mL In 20mg / mL bovine serum albumin BSA carbonate buffer solution (CBS, 0.1mol / L, pH9.6), magnetic stirring was carried out for...

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Abstract

The invention discloses an L-lauramide arginine monohydrochloride ethanol ester artificial antigen and a specific antibody as well as a preparation method and application thereof and belongs to the technical field of immunochemical analysis. The antigen is specially used for preparation of an L-lauramide arginine specific antibody and an enzyme-linked immunosorbent assay method, artificial antigens LAS-BSA and LAS-OVA are synthesized, an immune rabbit obtains an L-lauramide arginine specific polyclonal antibody, and finally an enzyme-linked Immunoassay method is established by utilizing the synthesized coating antigen and the specific antibody. The method has the lowest LAE detection limit of 0.005mg / L, recovery is 82.4-109.8%, the average variation coefficient is 3.2-5.8%, and the method has the characteristics of high accuracy and high sensitivity and accords with residue analysis standard.

Description

technical field [0001] The invention belongs to the technical field of immunochemical analysis, and in particular relates to a novel food preservative L-lauramide arginine hydrochloride ethanol ester, a preparation method and application of an antigen and a specific antibody. Background technique [0002] L-lauroylamide arginine hydrochloride ethanol ester (ethyl lauroyl arginate (LAE)), the molecular formula is C 20 h 41 ClN 4 o 3 , with the following structure: [0003] [0004] The appearance of LAE is white powder, its solubility in water at 20°C is 247g / kg, its chemical properties are stable in the range of pH3-7, it has high antibacterial activity, its melting point is 50-58°C, and its distribution coefficient in water and oil greater than 10. That is, it mainly exists in the water phase. Compared with other preservatives with different chemical structures and similar functions, LAE can be more conveniently dissolved in water-based food and cosmetic systems, an...

Claims

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Application Information

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IPC IPC(8): C07C279/14C07K14/765C07K14/77C07K1/107C07K16/44G01N33/543G01N33/531
CPCG01N33/531G01N33/543C07C279/14C07K14/765C07K14/77C07K16/44C07K19/00
Inventor 付岩张艳张亮陈国吴银良
Owner NINGBO ACAD OF AGRI SCI
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