Preparation method of perfluorooctanoic acid artificial complete antigen
A technology of perfluorooctanoic acid and complete antigen, applied in chemical instruments and methods, animal/human proteins, instruments, etc.
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Embodiment 1
[0006]
[0007] Perfluorooctanoic acid (10mg), NHS (14mg), EDC (40mg) were dissolved in DMF (3ml), under nitrogen protection, reacted at room temperature for 10 hours; then the above reaction mixture was centrifuged, and the supernatant was slowly added dropwise to BSA ( 40mg) in PBS buffer solution (20ml). React at room temperature for 5 hours, then centrifuge, put the supernatant into a dialysis bag prepared by a dialysis membrane with a molecular weight of 10,000, suspend the dialysis bag in PBS buffer solution in a 2000ml beaker, dialyze at 4°C for 7 days, and then put the dialysis bag The liquid freeze-dried in is the complete antigen of synthetic perfluorooctanoic acid. The molecular weight of the antigen determined by MALDI-TOF-MS mass spectrometry is: 72593.
Embodiment 2
[0009]
[0010] Dissolve perfluorooctanoic acid (100mg), compound A (40mg), and compound B (0.05ml) in NMM (3ml), under nitrogen protection, react at -20°C for 5 hours, perform conventional organic reaction treatment, and obtain compound C after column chromatography Pure. Compound C (200 mg) and compound D (230 mg) were dissolved in pyridine, DMAP (250 mg) was added at 0°C, reacted for 5 hours, conventional organic reaction treatment, and purified compound E was obtained after column chromatography. Compound E (40 mg) was dissolved in DMF (2 ml), and then slowly added dropwise to a PBS buffer solution (20 ml) of BSA (140 mg). React at room temperature for 5 hours, then centrifuge, put the supernatant into a dialysis bag prepared by a dialysis membrane with a molecular weight of 10,000, suspend the dialysis bag in PBS buffer solution in a 2000ml beaker, dialyze at 4°C for 7 days, and then put the dialysis bag The liquid freeze-dried in is the complete antigen of synthetic ...
Embodiment 3
[0012]
[0013] Dissolve perfluorooctanoic acid (100mg), compound A (40mg), and compound B (0.05ml) in NMM (3ml), under nitrogen protection, react at -20°C for 5 hours, perform conventional organic reaction treatment, and obtain compound C after column chromatography Pure. Compound C (200 mg) and compound D (230 mg) were dissolved in DCM, DMAP (250 mg) was added at 0°C, reacted for 5 hours, conventional organic reaction treatment, and purified compound E was obtained after column chromatography. Compound E (40 mg) was dissolved in DMF (2 ml), and then slowly added dropwise to a PBS buffer solution (20 ml) of BSA (140 mg). React at room temperature for 5 hours, then centrifuge, put the supernatant into a dialysis bag prepared by a dialysis membrane with a molecular weight of 10,000, suspend the dialysis bag in PBS buffer solution in a 2000ml beaker, dialyze at 4°C for 7 days, and then put the dialysis bag The liquid freeze-dried in is the complete antigen of synthetic perfl...
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