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
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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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