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Ethyl sulfate artificial antigen and preparation method and application thereof

An ethyl sulfate, artificial antigen technology, applied in the preparation of sulfate, the preparation method of peptide, chemical instruments and methods, etc. problem, to achieve the effect of high accuracy, high specificity and specificity, and accurate test results

Active Publication Date: 2020-08-11
HANGZHOU BIOTEST BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are many problems with the breath alcohol detector, which affect the accuracy of the alcohol test results
Because there are many uncertain factors affecting the test results of the breath alcohol detector, for example, the breath alcohol detection technology requires the collection of the gas exhaled from the deep lungs of the human body, so the way of exhalation, the length of time, and the air flow rate The size and the degree of diffusion of exhaled breath in the air will affect the accuracy of the test; for example, the breath alcohol detector needs to exhale deeply for 2.5 seconds, which is difficult to master in actual operation; for example, before the breath test, Hyperventilation or high-frequency deep breathing by the driver will reduce the alcohol concentration in the exhaled gas by 20%. If you hold your breath for 20 seconds before the test, the alcohol concentration in the exhaled gas will increase by 15%. Therefore, the driver Breathing patterns can also affect the accuracy of test results

Method used

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  • Ethyl sulfate artificial antigen and preparation method and application thereof
  • Ethyl sulfate artificial antigen and preparation method and application thereof
  • Ethyl sulfate artificial antigen and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] (1) Preparation of ethylsulfate hapten

[0037] Weigh 100mg (0.71mmol) 2-aminoethanol hydrogen sulfate, join in 50ml one-necked round bottom flask, then add 5mlN, N-dimethylformamide (DMF), 276mg (2mmol) potassium carbonate and 300mg (1.31 mmol) 2-(4-(bromomethyl)phenyl)acetic acid, add a stirring bar, and stir the reaction at room temperature for 20 hours; Dichloromethane:methanol=1:1 (volume ratio), product ratio shift value Rf=0.3~0.4; obtain hapten (2) 86mg, specific synthetic route is as follows figure 1 shown.

[0038] (2) Preparation of ethyl sulfate artificial antigen

[0039] a. Weigh 50mg (0.17mmol) ethyl sulfate hapten in a 50ml round bottom flask, add 2.5ml N,N-dimethylformamide (DMF), then add 39mg (0.34mmol) N-hydroxysuccinyl Imine (NHS) and 70 mg (0.34 mmol) N,N-dicyclohexylcarbodiimide (DCC) were stirred at room temperature overnight, centrifuged after the reaction, and the supernatant was taken as solution A.

[0040] b. Weigh 14.5 g of disodium hyd...

Embodiment 2

[0053] (1) Preparation of ethylsulfate hapten

[0054] Weigh 100mg (0.71mmol) of 2-aminoethanol hydrogen sulfate and add it to a 50ml single-necked round bottom flask, then add 5mlN,N-dimethylformamide (DMF), 138mg (1mmol) potassium carbonate and 200mg (0.87 mmol) 2-(4-(bromomethyl)phenyl)acetic acid, add a stirring bar, and stir the reaction at room temperature for 20 hours; Dichloromethane:methanol=1:1 (volume ratio), product ratio shift value Rf=0.3~0.4; obtain 40mg of hapten (2), the specific synthetic route is as follows figure 1 shown.

[0055] (2) Preparation of ethyl sulfate artificial antigen

[0056] a. Weigh 40mg (0.14mmol) ethyl sulfate hapten in a 50ml round bottom flask, add 2ml N,N-dimethylformamide (DMF), then add 24mg (0.21mmol) N-hydroxysuccinyl Amine (NHS) and 43mg (0.21mmol) N,N-dicyclohexylcarbodiimide (DCC), stirred at room temperature overnight, centrifuged after the reaction, and the supernatant was taken as solution A.

[0057] b. Weigh 14.5 g of d...

Embodiment 3

[0063] (1) Preparation of ethylsulfate hapten

[0064] Weigh 100mg (0.71mmol) 2-aminoethanol hydrogen sulfate, join in 100ml single-necked round bottom flask, then add 10mlN,N-dimethylformamide (DMF), 552mg (4mmol) potassium carbonate and 600mg (2.62 mmol) 2-(4-(bromomethyl)phenyl)acetic acid, add a stirring bar, and stir the reaction at room temperature for 20 hours; Dichloromethane:methanol=1:1 (volume ratio), product ratio shift value Rf=0.3~0.4; obtain hapten (2), the specific synthetic route is as follows figure 1 shown.

[0065] (2) Preparation of ethyl sulfate artificial antigen

[0066] a. Weigh 50mg (0.17mmol) of ethyl sulfate hapten in a 100ml round bottom flask, add 10ml of N,N-dimethylformamide (DMF), then add 1.1mmol of N-hydroxysuccinimide (NHS) and 1.0 mmol N,N-dicyclohexylcarbodiimide (DCC), stirred at room temperature overnight, centrifuged after the reaction, and the supernatant was taken as liquid A.

[0067] b. Weigh 14.5 g of disodium hydrogen phosphat...

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Abstract

The invention discloses an ethyl sulfate artificial antigen and a preparation method and application thereof. According to the invention, 2-aminoethanol hydrogen sulfate is used as a raw material to prepare the ethyl sulfate artificial antigen; a 2-(4-(bromomethyl) phenyl) acetic acid connecting arm is introduced into amino to prepare an ethyl sulfate hapten, the hapten reacts with N-hydroxysuccinimide and N, N-cyclohexylcarbodiimide to obtain an active ester, and a coupling reaction is performed on the active ester and protein to obtain the ethyl sulfate artificial antigen. According to the invention, the hapten can be obtained by only one step, the process is simple, the raw material source is not limited, the characteristic structure of ETS is also retained, and the specificity is verygood; the connecting arm introduced by the invention can play the immunogenicity of the antigen to a greater extent, enhance the immune effect and stimulate animals to generate antibodies, and can effectively make up the defect that ETS is difficult to stimulate organisms to generate antibodies due to the execessively simple structure of ETS.

Description

technical field [0001] The invention specifically relates to an ethyl sulfate artificial antigen, a preparation method and an application. Background technique [0002] Ethanol, commonly known as alcohol, is one of the most abused substances worldwide. Statistics show that drivers who drink and drive are 15 times more likely to have an accident than usual, and 30% of road traffic accidents are caused by drunk driving or drunk driving. 59 percent of driver deaths on file were related to DUI. Shocking figures tell us that drunk driving seriously endangers traffic safety, harms others and harms society. [0003] At present, the traffic police usually use a breath alcohol detector to test the alcohol of the driver. However, there are many problems in the breath alcohol detector, which affect the accuracy of the alcohol test results. Because there are many uncertain factors affecting the test results of the breath alcohol detector, for example, the breath alcohol detection te...

Claims

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Application Information

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IPC IPC(8): C07C305/04C07C303/24C07K14/765C07K1/107C07K16/44G01N33/577G01N33/558G01N33/53
CPCC07C305/04C07K14/765C07K16/44G01N33/577G01N33/558G01N33/5308Y02P20/55
Inventor 王百龙王晨苏祥煌
Owner HANGZHOU BIOTEST BIOTECH CO LTD
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