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Chlorpromazine hapten, artificial antigen, antibody as well as preparation method and application thereof

An artificial antigen, chlorpromazine technology, applied in chemical instruments and methods, animal/human proteins, serum albumin, etc., can solve the problems of insufficient specificity and recognition of chlorpromazine antibodies, lack of chlorpromazine, etc. Accurate, specific, and sensitive effects

Active Publication Date: 2011-03-23
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Immunological analysis technology has been widely used in the field of drug residue detection due to its advantages of sensitivity, specificity, rapidity and simplicity. The synthesis of artificial antigen is the key to the establishment of hapten substance immunoassay method. For chlorpromazine, because the hapten design and synthetic reasons, the specificity and recognition of the chlorpromazine antibody obtained by the prior art are not enough, and the relevant technical reports of the immunological analysis method of chlorpromazine are lacking

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  • Chlorpromazine hapten, artificial antigen, antibody as well as preparation method and application thereof
  • Chlorpromazine hapten, artificial antigen, antibody as well as preparation method and application thereof
  • Chlorpromazine hapten, artificial antigen, antibody as well as preparation method and application thereof

Examples

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Embodiment 2

[0044] Example 2: Synthesis and identification of artificial antigens

[0045] 1. Synthesis and identification of chlorpromazine antigen (III)

[0046] Chlorpromazine antigen (III) glutaraldehyde synthesis method:

[0047] (1) Weigh 35 mg of chlorpromazine hapten and 68 mg of BSA and dissolve them in 10 mL of phosphate buffered saline (PBS) solution, and stir well at room temperature;

[0048] (2) Drop 40 μL of glutaraldehyde into the mixed solution described in step (1), and after the addition is complete, stir and react at room temperature for 3 hours;

[0049] (3) The solution after the reaction in step (2) is packed in a dialysis bag, and dialyzed with PBS solution at 4°C for 72 hours, during which the dialysate is changed 6 times, to obtain the target product, which is aliquoted and stored at -20°C.

[0050] Human serum protein, hemocyanin or ovalbumin can also be used in the above method. The preparation method of chlorpromazine-coated antigen is the same as the above method accord...

Embodiment 5

[0053] Example 5 Preparation and identification of chlorpromazine antigen

[0054] 2. Synthesis and identification of chlorpromazine antigen (II)

[0055] Diazotization synthesis of chlorpromazine antigen (II):

[0056] (1) Weigh 35mg of chlorpromazine hapten and dissolve it in 1mL of 1mol / L hydrochloric acid, adjust the pH of the solution to 2 with 1mol / L of hydrochloric acid;

[0057] (2) Take 1.1 mL of 1mol / L sodium nitrite solution and add it dropwise to the hapten solution within half an hour, and diazotize the solution at 0~5℃ for 1 hour;

[0058] (3) Weigh 68mg of BSA and dissolve it in 10mL PBS (phosphate buffered saline commonly used in laboratories) solution, add the solution after step (2) to the BSA-PBS solution dropwise, and use 1mol / L sodium hydroxide Adjust the pH of the solution to 7.5, overnight at 4°C;

[0059] (4) The solution after the reaction in step (3) is packed in a dialysis bag, and dialyzed with PBS solution at 4°C for 72 hours, during which the dialysate is c...

Embodiment 3

[0062] Example 3: Preparation of monoclonal, polyclonal antibodies or chlorpromazine genetic engineering antibodies

[0063] (1) Preparation of monoclonal antibodies

[0064] Animal immunization: Balb / c mice were immunized with a hapten containing a 2-carbon carboxyl spacer and a carrier protein conjugate as an immunogen. The indirect immunoassay was tested and the blood contained chlorpromazine specific antibodies. Mouse spleen.

[0065] Cell fusion and cloning: The spleen cells of Balb / c mice that produce specific antibodies are fused with myeloma cells SP20, and the cell supernatant is determined by indirect competitive time-resolved immunoassay to screen positive wells. The positive wells are cloned by limiting dilution method or microscopic cloning method to obtain and establish hybridoma cell lines producing monoclonal antibodies.

[0066] Cell cryopreservation and resuscitation: Take the hybridoma cells in the logarithmic growth phase to make a cell suspension with cryopreserv...

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Abstract

The invention provides a chlorpromazine hapten, an artificial antigen and an antibody as well as a preparation method and application thereof. The chlorpromazine artificial hapten has a molecular structure (I); the chlorpromazine artificial antigen has molecular structures (II) or (III); the antiserum obtained by the invention has the titer of 1:3*105, the lowest detection limit of 3ng / mL and the half-inhibition concentration of 29ng / mL; and the generated antibody has high specificity, sensitivity and accuracy. The antigen and the antibody have wide application prospect in the detection of residual chlorpromazine in the food.

Description

technical field [0001] The invention belongs to the technical field of food safety. It specifically relates to the preparation method and application of chlorpromazine hapten, artificial antigen and antibody. Background technique [0002] Chlorpromazine (Chlorpromazine, CPZ) is also known as Dongmianling, and its molecular formula is: C 17 h 19 C 1 N 2 S, generally in the form of hydrochloride for medicinal use, the chemical name is: N, N-dimethyl-2-chloro-10H-phenothiazine-10-propylamine hydrochloride. Belonging to one of the phenothiazine drugs, it is a central dopamine receptor blocker with sedative, antipsychotic, antiemetic, hypothermic and basal metabolism, -Adrenergic receptor and m-cholinergic receptor blockade, antihistamine, endocrine effects, etc. It is clinically used to control symptoms such as restlessness, nervousness, hallucinations, and delusions of schizophrenia or other mental illnesses; to treat vomiting caused by various reasons; it is also used f...

Claims

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Application Information

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IPC IPC(8): C07D279/34C07K14/765C07K14/77C07K14/795C07K16/44G01N33/53G01N33/566G01N33/577
Inventor 孙远明沈玉栋徐振林王弘肖犇雷红涛杨金易
Owner SOUTH CHINA AGRI UNIV
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