Baclofen artificial hapten, artificial antigen as well as 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, etc., can solve the problems of artificial haptens and artificial antigens without baclofen, and achieve accurate immune detection And immunoassay, high sensitivity, strong specificity effect

Active Publication Date: 2020-10-30
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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  • Baclofen artificial hapten, artificial antigen as well as preparation method and application thereof
  • Baclofen artificial hapten, artificial antigen as well as preparation method and application thereof
  • Baclofen artificial hapten, artificial antigen as well as 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 is as follows: figure 1 ), including the following steps:

[0052] (1) Preparation of artificial hapten:

[0053] ① Weigh 200mg (0.94mmol) of baclofen into a 50ml single-necked round-bottom single-necked flask, add 10ml of benzene dried by 3A molecular sieves, install a condenser tube and a drying tube, add a stirring bar and stir in an ice-water bath at 0°C for 5 minutes, Then add 143 μL (1.03 mmol) of trifluoroacetic anhydride, remove the ice-water bath, stir and react at room temperature for 1 hour, then transfer to 80°C oil bath and continue to stir and reflux for 3 hours; finish the reaction, cool to room temperature, take out the rotor, and decompress The solvent was evaporated to give 262mg (0.85mmol) of light yellow oily substance A;

[0054] The light yellow oil A was subjected to TLC detection, the chromatographic solution was ethyl acetate:petroleum ether=10:3 (v / v...

Embodiment 2

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

[0125] (1) Identification of baclofen artificial antigen:

[0126] Molar absorption coefficient ε: Prepare baclofen artificial hapten solutions with concentrations of 0 μg / ml, 5 μg / ml, 10 μg / ml, 20 μg / ml, 30 μg / ml, and 40 μg / ml in PBS buffer, and the baclofen artificial hapten solution can be known from the ultraviolet scanning diagram The maximum absorption wavelength of the chlorofen hapten is 245nm, and the absorbance value is measured at 245nm with PBS as a control sample, and parallel samples are made for each concentration. The formula for calculating the molar absorptivity (ie, the molar absorptivity) is: ε=absorbance value / molar concentration.

[0127] Determination of conjugate protein concentration: Prepare the concentration of 0μg / ml, 10μg / ml, 20μg / ml, 30μg / ml, 40μg / ml, 60μg / ml, 80μg / ml, 100μg / ml, 120μg / ml in PBS buffer Add 3ml of Coomassie Brilliant Blue staining solution to 1ml of b...

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Abstract

The invention discloses a baclofen artificial hapten, an artificial antigen as well as a preparation method and application thereof. The molecular structural formula of the baclofen artificial haptenis represented by a formula (I), and the molecular structural formula of the baclofen artificial antigen is represented by a formula (II). According to the baclofen artificial hapten, the characteristic structure of baclofen is reserved to the maximum extent, and the baclofen artificial hapten has an active group capable of being coupled with carrier protein and can be used as an antigen determinant; the further prepared baclofen artificial antigen can be used for immunizing to obtain an anti-baclofen antibody with high affinity, high sensitivity and strong specificity, the titer of immune serum obtained by immunizing New Zealand white rabbits is up to 1:80000, and the antibody can be used for carrying out rapid and accurate immunodetection and immunoassay on baclofen.

Description

technical field [0001] The invention belongs to the technical field of biochemical engineering, and in particular 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 spasm symptoms of increased muscle tone caused by pyramidal tract damage, spastic hemiplegia and paraplegia caused by different reasons, such as multiple sclerosis, cerebrovascular disease, spinal cord injury and sequelae of myelitis , children with cerebral palsy, tetanus, refractory hiccups, etc. [0003] The drug is rapidly and completely absorbed from the gastrointestinal tract after oral administration. 1 to 3 hours after oral administration, the plasma concentration reaches the peak, but individual differences are large. 70% to 80% of the drug is excreted in the urine in its original form, and about 15% is metabolized by the liver. [0004] Long-term use ...

Claims

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Application Information

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Patent Type & Authority Applications(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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