Paraquat hapten, paraquat complete antigen and preparation method of paraquat hapten and paraquat complete antigen
A technology of complete antigen and paraquat, which is applied to the preparation method of peptides, chemical instruments and methods, animal/human proteins, etc., can solve the problems of complex processing, cumbersome operation, and poor measurement accuracy, and achieve simple and effective synthesis steps. Good specificity and low cost effect
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Embodiment 1
[0032] Example 1, Preparation and identification of paraquat hapten
[0033] 1. Preparation of Paraquat Hapten
[0034] The synthesis route of paraquat hapten is as follows:
[0035]
[0036] Specific synthesis process:
[0037] (1) Synthesis of N-methyl-dipyridine iodide A 250ml three-necked round-bottomed flask was fixed on an iron stand, protected from light and passed through nitrogen. 808 mg of 4,4'-bipyridine was dissolved in 8 ml of acetone, stirred and cooled to 4°C, and 400 μl of methyl iodide was added dropwise to the above solution. Keep stirring at 4°C. The end point of the reaction was controlled by TLC (developing solvent: methanol: glacial acetic acid = 5:2). The reaction reached equilibrium in about 6 hours. Filter, wash with acetone, and drain to obtain yellow needle-like crystals. The crystals were recrystallized from absolute ethanol. Vacuum dry.
[0038] (2) Synthesis of N-methyl-N'-pentanoic acid ethyl ester group-dipyridine iodide bromide 0.85m...
Embodiment 2
[0042] Example 2, Preparation and Identification of Paraquat Complete Antigen
[0043] 1. Preparation of paraquat complete antigen
[0044] The synthetic route of paraquat complete antigen is as follows:
[0045]
[0046]Specific synthesis process: Dissolve 9 mg of paraquat hapten in 500 ul of dimethyl sulfoxide (DMSO), add 15 ul of tri-n-butylamine, stir at room temperature for 5 min, then add 8 ul of isobutyl chloroformate, and stir at room temperature for 30 min. 15 mg of BSA was dissolved in 2 ml of borate buffer solution pH 8.7. Slowly drop the paraquat hapten solution into the BSA solution. Stir overnight at 4°C, dialyze with PBS for three days, and change the dialysate every four hours. Aliquot into EP tubes and store at -20°C.
[0047] 2. Identification of paraquat complete antigen
[0048] According to the increase of the molecular weight of the complete antigen after coupling compared to the molecular weight of the carrier protein, the coupling result was ide...
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