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.

Inactive Publication Date: 2011-12-14
RES CENT FOR ECO ENVIRONMENTAL SCI THE CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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  • Preparation method of perfluorooctanoic acid artificial complete antigen
  • Preparation method of perfluorooctanoic acid artificial complete antigen
  • Preparation method of perfluorooctanoic acid artificial complete antigen

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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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Abstract

The invention relates to a preparation method of perfluorooctanoic acid (Perfluorooctanoic acid, PFOA) artificial complete antigen, which belongs to the technical field of organic chemistry and immunology. The carboxylic acid group of perfluorooctanoic acid is used to link perfluorooctanoic acid to a carrier protein directly or through a connecting arm of an appropriate length to prepare a complete antigen of perfluorooctanoic acid. The carrier protein can be bovine serum albumin (BSA), ovalbumin (OVA), hemocyanin (KLH) or human serum albumin (HSA). Determination of the amount of PFOA on the carrier protein by MALDI-TOF-MS mass spectrometry. The method is simple to operate, does not require special instruments and equipment, and has low cost, which provides a basis for screening its specific antibody and establishing an immunoassay method in the future.

Description

technical field [0001] The invention relates to a preparation method of perfluorooctanoic acid (Perfluorooctanoic acid, PFOA) artificial complete antigen, which belongs to the technical field of organic chemistry and immunology. Background technique [0002] Perfluorooctanoic acid (PFOA) is an important fluorine-containing surfactant, which is mainly used in polymer additives, lubricants, fire extinguishing agents, insecticides, etc. Due to the chemical inertness of perfluorooctanoic acid and its extensive production and use, it has caused serious environmental accumulation and pollution, and has become a new type of persistent organic pollutant that has attracted increasing attention after organochlorine pesticides and dioxins. Studies have shown that perfluorooctanoic acid is a liver carcinogen with moderate toxicity, which can increase the risk of human cancer. In addition, perfluorooctanoic acid has embryotoxicity and potential neurotoxicity, and is highly bioaccumulativ...

Claims

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Application Information

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IPC IPC(8): C07K14/765C07K14/77C07K14/795G01N27/62
Inventor 魏国华孙艳灵杜宇国
Owner RES CENT FOR ECO ENVIRONMENTAL SCI THE CHINESE ACAD OF SCI
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