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Gemfibrozil hapten, artificial antigen and antibody and preparation method thereof and purpose thereof

A technology of gemfibrozil and artificial antigen, which is applied in the fields of peptide preparation, chemical instruments and methods, carboxylate preparation, etc. The effect of good affinity and low cross-reaction rate

Active Publication Date: 2019-01-18
BEIJING KWINBON BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The instrumental detection method has the disadvantages of cumbersome sample pretreatment, long detection time, and expensive instruments, so it cannot be widely used in my country, and it does not meet the requirements of on-site detection "accurate detection and screening of a large number of samples in a short period of time and at low cost".

Method used

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  • Gemfibrozil hapten, artificial antigen and antibody and preparation method thereof and purpose thereof
  • Gemfibrozil hapten, artificial antigen and antibody and preparation method thereof and purpose thereof
  • Gemfibrozil hapten, artificial antigen and antibody and preparation method thereof and purpose thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1: Hapten Synthesis

[0025] A gemfibrozil hapten, the molecular structural formula is:

[0026]

[0027] It is used as the raw material of the antigen system for animal immunization.

[0028] The preparation method of above-mentioned gemfibrozil hapten is as follows (synthetic route sees attached figure 1 ):

[0029] Take 0.5g of 2-hydroxy-4-methylbenzaldehyde, add 50mL of acetonitrile to dissolve, clarify, add 0.61g of potassium carbonate, add 0.84g of 5-bromo-2,2-dimethylpentanoic acid, heat under reflux in oil bath for 3h ; Stop the reaction, cool to room temperature, rotary evaporate, remove acetonitrile, add water 50mL, ethyl acetate 100mL×3, extract three times, combine the organic phases, put on a silica gel column, and use ethyl acetate-petroleum ether chromatography with a volume ratio of 1:3 After purification, the gemfibrozil hapten can be obtained.

Embodiment 2

[0030] Example 2: Artificial Antigen Synthesis and Identification

[0031] A kind of gemfibrozil artificial antigen made by the gemfibrozil hapten described in embodiment 1, molecular structural formula is:

[0032]

[0033] 1. Synthesis of immunogen

[0034] The immunogen was synthesized as follows:

[0035] Take 9.8 mg of gemfibrozil hapten, add 0.2 mL of ethanol to dissolve to obtain liquid A; take 50 mg of carrier protein, add 3 mL of 0.1 mol / L sodium bicarbonate solution to dissolve to obtain liquid B; add liquid A dropwise to liquid B , stirred at 4°C for 24h, purified by dialysis with normal saline for 3d, changed the solution 3 times a day, aliquoted, and stored at -20°C for later use.

[0036] Identification of artificial antigens:

[0037] According to the ratio of hapten, carrier protein and coupling product used in the synthetic gemfibrozil immunogen reaction, carry out ultraviolet (200-400nm) scanning measurement, and calculate the binding ratio by comparing...

Embodiment 3

[0042] Example 3: Preparation of Monoclonal Antibody

[0043] A gemfibrozil-specific antibody is the monoclonal immunoglobulin G obtained by immunizing white mice with the gemfibrozil immunogen described in Example 2, and can have a specific immune reaction with gemfibrozil, used for Detection of gemfibrozil residues.

[0044] The preparation method of above-mentioned gemfibrozil monoclonal antibody is as follows:

[0045] 1) Animal immunization: the immunogen was injected into the body of Balb / c mice, and the immunization dose was 150 μg / mouse to make them produce antiserum.

[0046] 2) Cell fusion and cloning: the splenocytes of Balb / c mice producing specific antibodies were fused with myeloma cells SP20, the cell supernatant was measured by an indirect competitive enzyme-linked immunoassay method, and positive wells were screened. Positive wells were cloned by limiting dilution method to obtain and establish monoclonal antibody-producing hybridoma cell lines.

[0047]3) ...

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Abstract

The invention provides a gemfibrozil hapten and an artificial antigen. Molecular structural formulas of the gemfibrozil hapten and the artificial antigen are correspondingly as shown in the description. The invention further discloses preparation methods for the gemfibrozil hapten, the artificial antigen and an antibody prepared from the gemfibrozil hapten and the artificial antigen and purposes of the gemfibrozil hapten and the antibody. The antibody which is finally prepared by the preparation method provided by the invention has the advantages of high detection sensitivity and high specificity.

Description

technical field [0001] The present invention relates to selecting a compound having -CHO and possibly containing the original structure of gemfibrozil as the gemfibrozil hapten, artificial antigens and specific antibodies made of this hapten; and such hapten Uses of antigens, artificial antigens, and specific antibodies, and methods for preparing haptens, artificial antigens, and antibodies. Background technique [0002] Pharmaceuticals and personal care products (PPCPs) are the necessities of human beings, and there are various types, including various antibiotics, synthetic musk, painkillers, antihypertensive drugs, contraceptives, hypnotics, weight loss drugs, hair sprays, hair dyes and fungicides, etc. Gemfibrozil (GEM) is a lipid-regulating drug used to treat diseases such as hyperlipidemia. GEM cannot be completely removed after entering the sewage treatment plant, and can enter the water environment with the effluent, causing secondary pollution. Because of its low ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07C59/74C07C51/367G01N33/577G01N33/94C07K14/765C07K14/77C07K1/107C07K16/44
CPCG01N33/577G01N33/94C07C59/74C07K14/765C07K14/77C07K16/44C07K19/00
Inventor 万宇平何方洋赵正苗张瑜韩深崔廷婷崔海峰王兆芹杨春艳丛倩千
Owner BEIJING KWINBON BIOTECH
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