A kind of baclofen artificial hapten, artificial antigen and preparation method and application thereof

A technology of artificial haptens and artificial antigens, which is applied in the preparation of carboxylic acid amides, chemical instruments and methods, and the preparation of organic compounds, can solve the problems of artificial haptens and artificial antigens without baclofen, and achieve accurate immune detection And immunoassay, high affinity, high sensitivity effect

Active Publication Date: 2022-07-05
HANGZHOU TONGZHOU BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In the prior art, there are no related reports of baclofen artificial hapten and artificial antigen

Method used

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  • A kind of baclofen artificial hapten, artificial antigen and preparation method and application thereof
  • A kind of baclofen artificial hapten, artificial antigen and preparation method and application thereof
  • A kind of baclofen artificial hapten, artificial antigen and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] This implements a kind of preparation method of baclofen artificial antigen (reaction process such as figure 1 ), including the following steps:

[0052] (1) Preparation of artificial hapten:

[0053] ①Weigh 200mg (0.94mmol) of baclofen into a 50ml single-necked round-bottomed single-necked flask, add 10ml of benzene dried by 3A molecular sieves, install a condenser tube and a drying tube, add a stirrer and stir in an ice-water bath at 0°C for 5 minutes, Then 143 μL (1.03 mmol) of trifluoroacetic anhydride was added, the ice-water bath was removed, and the reaction was stirred at room temperature for 1 hour, then transferred to an oil bath at 80° C. to continue stirring and refluxing for 3 hours; the reaction was completed, cooled to room temperature, the rotor was taken out, and the pressure was reduced. The solvent was evaporated to dryness to obtain a pale yellow oil A262mg (0.85mmol);

[0054] The light yellow oil A was detected by TLC, the chromatographic liquid ...

Embodiment 2

[0124] The performance measurement of embodiment 2 baclofen artificial antigen

[0125] (1) Identification of Baclofen Artificial Antigen:

[0126] Molar absorption coefficient ε: Baclofen artificial hapten solution with concentrations of 0μg / ml, 5μg / ml, 10μg / ml, 20μg / ml, 30μg / ml and 40μg / ml was prepared in PBS buffer. The maximum absorption wavelength of chlorofen hapten was 245nm, and the absorbance value was measured at 245nm with PBS as a control sample, and parallel samples were made for each concentration. The formula for calculating the molar absorption coefficient (ie, the molar absorption coefficient) is: ε=absorbance value / molar concentration.

[0127] Determination of conjugate protein concentration: PBS buffer was used to prepare the concentration of 0μg / ml, 10μg / ml, 20μg / ml, 30μg / ml, 40μg / ml, 60μg / ml, 80μg / ml, 100μg / ml, 120μg / ml 1ml each of the bovine serum albumin solution, add 3ml Coomassie brilliant blue staining solution, mix immediately, warm in a water bat...

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Abstract

The invention discloses a baclofen artificial hapten, an artificial antigen and a preparation method and application thereof. The molecular structural formula of the baclofen artificial hapten is as formula (I), and the molecular structural formula of the baclofen artificial antigen is as formula (II). The baclofen artificial hapten of the present invention retains the characteristic structure of baclofen to the greatest extent, and has an active group that can be coupled with a carrier protein, and can be used as an antigenic determinant; the baclofen artificial antigen obtained by further preparation Anti-baclofen antibodies with high affinity, high sensitivity and specificity can be obtained by immunization. The titer of the immune serum obtained by immunizing New Zealand white rabbits is as high as 1:80,000, which can be used for rapid and accurate immunological detection and detection of baclofen. Immunoassay.

Description

technical field [0001] The invention belongs to the technical field of biochemical industry, and particularly relates to a baclofen artificial hapten, an artificial antigen and a preparation method and application thereof. Background technique [0002] Baclofen is a skeletal muscle relaxant used to improve the spastic symptoms of increased muscle tone caused by damage to the pyramidal tract, spastic hemiplegia and paraplegia caused by different causes, such as multiple sclerosis, cerebrovascular disease, spinal cord injury and myelitis sequelae , children with cerebral palsy, tetanus, refractory hiccups, etc. [0003] This drug is rapidly and completely absorbed from the gastrointestinal tract after oral administration. 1 to 3 hours after oral administration, the plasma concentration reaches a peak, but individual differences are large. 70% to 80% of the drug is excreted from the urine in its original form, and about 15% is metabolized by the liver. [0004] Sudden discon...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07C237/22C07C231/14C07K14/765C07K16/06C07K16/44G01N33/53
CPCC07C237/22C07K14/765C07K16/44C07K16/06G01N33/94Y02P20/55
Inventor 王镇邵越水
Owner HANGZHOU TONGZHOU BIOTECHNOLOGY CO LTD
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