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Preparation of semicarbazide derivative hapten, antigen and antibody

A semicarbazide and derivative technology, applied in the field of immunochemistry, can solve the problems that there are no technical reports on semicarbazide derivative hapten, antigen and its specific antibody, and achieve high titer and good specificity

Inactive Publication Date: 2009-04-15
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, there are no technical reports on semicarbazide derivative hapten, antigen and its specific antibody

Method used

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  • Preparation of semicarbazide derivative hapten, antigen and antibody
  • Preparation of semicarbazide derivative hapten, antigen and antibody
  • Preparation of semicarbazide derivative hapten, antigen and antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Embodiment 1 The preparation method of semicarbazide derivative hapten MCPSEM

[0037] Add 1.15g of m-carboxybenzaldehyde into a 50ml round bottom flask, slowly add methanol until the p-carboxybenzaldehyde is completely dissolved, add 0.3g of semicarbazide during stirring, and stir overnight at room temperature; after the reaction is completed, filter, wash twice with water, and wash twice with methanol Repeatedly, dry, obtain 0.62g white powder, its structure is as shown in formula (I). The APCI-MS molecular ion peak of hapten MCPSEM is 207, HNMR (600MHz, d 6 -DMSO, TMS): δ10.35(s, 1H); 8.14(s, 1H); 8.03(d, J=7.8Hz, 1H); 7.93(s, 1H); 7.90(d, J=7.8Hz, 1H); 7.52 (t, J = 7.2 Hz, 1H,). The above spectral data can be assigned correctly, which is consistent with the structure of MCPSEM, indicating that the hapten MCPSEM was synthesized successfully.

[0038]

Embodiment 2

[0039] The preparation method of embodiment 2 semicarbazide derivative hapten CEPSEM

[0040] Add 1.06g of 2-(4-formylphenoxy)acetic acid into the round bottom flask, slowly add methanol until the 2-(4-formylphenoxy)acetic acid is completely dissolved, add 0.4g semicarbazide during stirring, and stir at room temperature After overnight, the reaction was completed, filtered, washed twice with water and twice with methanol, and dried to obtain 0.90 g of white powder, the structural formula of which was shown in formula (II). CEPSEM spectrum data: APCI-MSm / z: 237[M+H]+; 1H NMR (600MHz, d6-DMSO, TMS): δ10.09(s, 1H); 7.78(s, 1H); 7.64(d, J = 9.0 Hz, 2H); 6.91 (d, J = 9.0 Hz, 2H); 4.71 (s, 2H). The above spectral data can be assigned correctly, which is consistent with the CEPSEM structure, indicating that the hapten CEPSEM was synthesized successfully.

[0041]

Embodiment 3

[0042] The preparation method of embodiment 3 immunogen CEPSEM-BSA

[0043] Take the hapten CEPSEM 0.1mmol and dissolve it in 2ml DMF, add DCC 27.5mg and NHS 14.4mg with stirring. Magnetically stir the reaction overnight at 4°C. After centrifugation, the supernatant night is liquid A; weigh 140 mg of BSA and dissolve it in 10 ml of PBS (pH 8.0) with a concentration of 0.1 mol / L, add 1 ml of DMF, stir and dissolve to prepare liquid B; under magnetic stirring , solution A was gradually dropped into solution B, and reacted at 4°C for 12 hours. After centrifugation, the supernatant was taken and dialyzed with normal saline at 4°C for 3 days, and the dialysate was changed 3 times a day. The obtained whole antigen was dispensed into 0.5ml centrifuge tubes at a concentration of 1 mg / ml, and frozen in a -20°C refrigerator for immunization.

[0044] The prepared semicarbazide derivative immunogen has a structure shown in formula (V):

[0045]

[0046] Immunogen identification: The c...

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Abstract

The invention discloses a method for preparing semicarbazide derivative hapten, antigen and antibody; semicarbazide derivative hapten is obtained after semicarbazide is condensed with benzaldehyde ammonia aldehyde; the hapten is similar to the structure of semicarbazide derivative, thus showing up the veterinary drug molecular idiosyncratic antigenic determinant, and antigen can be prepared through an active ester method coupling with carrier protein; and by using the antigen immunization, laboratory animals can obtain antibody of high potency and good idiosyncracy aiming at semicarbazide derivative. The antibody can be used in rapidly detecting residual quantity of semicarbazide in animal-derived food, with lower limit of the detection up to 0.09 ng / mL.

Description

technical field [0001] The invention belongs to the technical field of immunochemistry, and in particular relates to a method for preparing a hapten, an antigen and an antibody of a semicarbazide derivative, and an application of the prepared hapten, antigen, antibody and antibody. Background technique [0002] Furacillin, as a nitrofuran drug, has been used to treat and prevent mammalian digestive tract diseases caused by Escherichia and Salmonella. Due to the carcinogenic effect of nitrofuran and its metabolite semicarbazide (SEM), the European Union banned the use of nitrofurans in food animals in 1995, and my country also issued a ban on the use of such veterinary drugs in 2002. At present, nitrofurazone is a mandatory inspection item in international animal-derived food trade, and has become a technical trade barrier for developed countries to restrict exports to third countries. Furacilin is sensitive to light, metabolizes rapidly, and exists in the form of metabolite...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07C281/10C07K16/06G01N33/536
Inventor 沈玉栋张世伟孙远明雷红涛王弘肖治理杨金易蔡肇婷
Owner SOUTH CHINA AGRI UNIV
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