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Sodium picosulfate hapten, artificial antigen and antibody and preparation method and application of same

A technology of sodium picosulfate and artificial antigen, applied in the field of antibody and its preparation, artificial antigen, sodium picosulfate hapten

Active Publication Date: 2021-01-22
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The key to the research and development of immunoassay methods is to design a suitable sodium picosulfate hapten and prepare antibodies with high sensitivity and specificity, but there is no information about sodium picosulfate hapten, artificial antigen, antibody related reports

Method used

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  • Sodium picosulfate hapten, artificial antigen and antibody and preparation method and application of same
  • Sodium picosulfate hapten, artificial antigen and antibody and preparation method and application of same
  • Sodium picosulfate hapten, artificial antigen and antibody and preparation method and application of same

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Experimental program
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Embodiment 1

[0104] Synthesis and identification of embodiment 1 sodium picosulfate hapten

[0105] Utilizing the structural characteristics of sodium picosulfate and on the basis of experimental data, two artificial haptens were designed.

[0106] 1. Synthesis and identification of the hapten 4-((4-(carboxymethoxy)phenyl)(pyridin-2-yl)methyl)phenylsulfate sodium (PC1)

[0107]

[0108] Concrete synthetic steps include:

[0109] Take 2-(4-((4-hydroxyphenyl)(pyridin-2-yl)methyl)phenoxy)ethyl acetate (1moL), triethylamine (4moL), with anhydrous dichloromethane as solvent , reacted with chlorosulfonic acid (1.5moL) under stirring at room temperature for 3-5h, separated and purified to obtain 4-((4-(2-ethoxy-2-oxyethoxy)phenyl)(pyridin-2-yl ) methyl) phenyl sulfuric acid. Dissolve 4-((4-(2-ethoxy-2-oxyethoxy)phenyl)(pyridin-2-yl)methyl)phenylsulfuric acid in methanol, 4-((4-(2- The volume ratio of ethoxy-2-oxyethoxy)phenyl)(pyridin-2-yl)methyl)phenylsulfuric acid to methanol is 1:1, add...

Embodiment 2

[0132] Synthesis and identification of embodiment 2 sodium picosulfate artificial antigen

[0133] 1. Synthesis of Sodium Picosulfate Artificial Antigen

[0134] The synthetic method of sodium picosulfate artificial antigen comprises the following steps:

[0135] Using PC1, PC2, PC3, and PC4 in Example 1 as haptens, respectively coupled lactoferrin (LF) and chicken ovalbumin (OVA) by active ester method, respectively weighed 1moL of the above-mentioned sodium picosulfate haptens, Dissolve 1.4moL NHS and 1.6moLEDC in 50-200μL DMF, and stir in the dark for 2-4 hours at room temperature to obtain sodium picosulfate hapten activation solution; add 10mg LF or OVA to 1mL PBS buffer (0.01moL / L , pH=7.4); slowly add the sodium picosulfate hapten activation solution dropwise into the LF solution, react at 4°C for 12 hours; dialyze with PBS buffer solution for 3 days, 3 times a day, and obtain sodium picosulfate after the dialysis Artificial antigen, aliquoted into centrifuge tubes, s...

Embodiment 3

[0147] The preparation of embodiment 3 sodium picosulfate antibody

[0148] Emulsify the prepared immunogen PC1-LF and immune adjuvant (incomplete Freund's adjuvant for the first immunization, and incomplete Freund's adjuvant for subsequent booster immunizations) in a volume ratio of 1:1, and immunize New Zealand white rabbit. Described New Zealand white rabbit body weight is 2.5~3kg, adopts neck and back subcutaneous multi-point injection, immunizes for the second time after 4 weeks, boosts immunization once every 3 weeks afterwards. One week after the third booster immunization, blood was collected from the ear vein, and the serum titer was determined by indirect competition ELISA. When the titer no longer rises, the marginal ear vein is used to boost the immunization. One week later, blood was collected from the heart, and the collected blood was obtained by warming for 0.5-1 hour at 37°C, then standing overnight at 4°C, and then sucking the precipitated serum with a stra...

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Abstract

The invention discloses sodium picosulfate hapten, artificial antigen and antibody and preparation method and application of same. The preparation method comprises the following steps: firstly, preparing sodium picosulfate hapten PC1, PC2, PC3 and PC4, coupling the hapten with carrier protein to obtain the artificial antigen, and immunizing animals to obtain the sodium picosulfate antibody; wherein preferably, the PC1-lactoferrin conjugate is used as an immunogen, the antibody obtained through immunization has high sensitivity and high specificity recognition capability on sodium picosulfate,IC50 is 5 ng / mL, and the cross reaction rate on structural analogues is lower than 10%. The invention also discloses a sodium picosulfate enzyme-linked immunoassay kit, a colloidal gold immunochromatographic assay kit and application of the sodium picosulfate enzyme-linked immunoassay kit and the colloidal gold immunochromatographic assay kit in food and / or health care products.

Description

technical field [0001] The present invention relates to the technical field of food and / or health product safety detection, more specifically, to a kind of sodium picosulfate hapten, artificial antigen, antibody and its preparation method and application. Background technique [0002] Sodium Picosulfate is a stimulant laxative that can treat constipation by stimulating intestinal peristalsis and secretion, and inhibiting water absorption in the intestinal cavity. Because of its remarkable laxative effect, some unscrupulous merchants add sodium picosulfate to enzyme foods or health foods claiming to have weight loss functions in order to achieve weight loss and bodybuilding effects. However, long-term consumption of excessive sodium picosulfate May cause disturbance of intestinal system or acute injury of gastric mucosa. [0003] At present, the most conventional detection method applied to sodium picosulfate in food is mainly high performance liquid chromatography-tandem ma...

Claims

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Application Information

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IPC IPC(8): C07D213/30C07K14/765C07K14/77C07K14/795C07K14/79C07K16/44G01N33/577G01N33/53
CPCC07D213/30C07K14/765C07K14/77C07K14/795C07K14/79C07K16/44G01N33/94G01N33/577
Inventor 雷红涛赖玮李向梅李兆栋沈兴徐振林沈玉栋杨金易孙远明
Owner SOUTH CHINA AGRI UNIV
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