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Prepn of specific antibody of provera acetate and method of using the antibody in homogenous or heterogenous enzyme-linked immune analysis

A technology of medroxyprogesterone acetate and chlormadinone acetate, applied in the field of establishment of ELISA analysis methods, can solve the problems of less research on ELISA methods, and achieve the effect of high sensitivity and strong specificity

Inactive Publication Date: 2007-11-07
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, there are few studies on the ELISA method of Medroxyprogesterone acetate residues at home and abroad, and the establishment of new antibodies and ELISA methods provides a powerful means for the analysis of Medroxyprogesterone acetate residues

Method used

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  • Prepn of specific antibody of provera acetate and method of using the antibody in homogenous or heterogenous enzyme-linked immune analysis
  • Prepn of specific antibody of provera acetate and method of using the antibody in homogenous or heterogenous enzyme-linked immune analysis
  • Prepn of specific antibody of provera acetate and method of using the antibody in homogenous or heterogenous enzyme-linked immune analysis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1: Synthesis of artificial hapten 3-carboxymethyloximino-6-methyl-17α-hydroxyl-pregnant-20-one acetate (3-CMO-MPA)

[0032] Reaction formula:

[0033]

[0034] According to the feeding ratio medroxyprogesterone acetate (MPA): carboxymethyloxamine hydrochloride=1: 1.2mol ratio, take by weighing 1mmol MPA and dissolve in methanol, then 1.2mmol carboxymethyloxamine hydrochloride and An appropriate amount of sodium acetate was dissolved therein to make the pH 6, and stirred at room temperature for 24 hours. After methanol was distilled off under reduced pressure, the residue was extracted with ethyl acetate (3×20ml), and then 10g Na 2 SO 4 Dry, filter, distill to remove ethyl acetate, and dry to obtain a white solid which is 3-CMO-MPA.

[0035] The above products were taken by ESI and 1 H-NMR determined its structure. The ESI molecular ion peak of this material is 459 (M - -1), 1 H-NMR (CDCl 3 ) is: δ0.66(s, 3H, 18-CH 3 ); 1.06 and 1.10 (2s, 3H, E and Z ...

Embodiment 2

[0036] Example 2: Synthesis of artificial hapten 3-carboxymethyloximino-6-chloro-17α-hydroxyl-pregn-6-en-20-one acetate (3-CMO-CMA)

[0037] Reaction formula:

[0038]

[0039] According to the feeding ratio of chlormadinone acetate: carboxymethyloxamine hydrochloride=1: 1.5mol ratio, take by weighing 1mmolCMA and dissolve in methanol, then mix 1.5mmol carboxymethyloxamine hydrochloride and an appropriate amount of sodium acetate Dissolve in it, make pH 6, stir at room temperature for 24 hours. After methanol was distilled off under reduced pressure, the residue was extracted with ethyl acetate (3×20ml), and then weighed with 10g Na 2 SO 4Dry, filter, distill to remove ethyl acetate, and dry to obtain a white solid which is 3-CMO-CMA.

[0040] The structures of the above products were determined by ESI and 1H-NMR respectively. The ESI molecular ion peak of this material is 477 (M - -1), 1 H-NMR (CDCl 3 ) is: δ0.70(s, 3H, 18-CH 3 ); 1.00 and 1.05 (2s, 3H, E and Z 19-...

Embodiment 3

[0041] Example 3: Synthesis of artificial hapten 3-carboxymethyloximino-6-methyl-17α-hydroxyl-pregn-6-en-20-one acetate (3-CMO-MEGA)

[0042] Reaction formula:

[0043]

[0044] According to the feeding ratio of megestrol acetate: carboxymethyl oxamine hydrochloride = 1: 1.5mol ratio, take 1mmol MEGA and dissolve it in methanol, then mix 1.5mmol carboxymethyl oxamine hydrochloride and appropriate amount of sodium acetate Dissolve in it, make pH 6, stir at room temperature for 24 hours. After methanol was distilled off under reduced pressure, the residue was extracted with ethyl acetate (3×20ml), and then weighed with 10g Na 2 SO4 was dried, filtered, ethyl acetate was distilled off, and dried to give a white solid as 3-CMO-MEGA.

[0045] The structures of the above products were determined by ESI and 1H-NMR respectively. The ESI molecular ion peak of this material is 477 (M - -1), 1 H-NMR (CDCl 3 ) is: δ0.70(s, 3H, 18-CH 3 ); 0.95 and 1.00 (2s, 3H, E and Z 19-CH 3 )...

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PUM

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Abstract

The present invention belongs to the field of enzyme-linked immunoassay technology, and is especially specific antibody of provera acetate and its preparation and usage in homogenous or heterogenous enzyme-linked immunoassay. The present invention prepares haptens 3-CMO-MPA, 3-CMO-CMA or 3-CMO-MEGA; couples hapten 3-CMO-MPA with protein to prepare immunological antigen; couples haptens 3-CMO-MPA, 3-CMO-CMA and 3-CMO-MEGA with protein separately to prepare three kinds of envelope antigens. When the immunological antigen is used in immunizing animal, the produced antibody can produce specific reaction with provera acetate and may be used in establishing homogenous or heterogenous enzyme-linked immunoassay method for detecting the content of provera acetate in the sample fast in high sensitivity.

Description

technical field [0001] Preparation of medroxyprogesterone acetate-specific antibody and method for using the antibody in homologous or heterologous enzyme-linked immunoassay, the present invention relates to an ELISA analysis method of medroxyprogesterone acetate with good specificity and high sensitivity The establishment of the method belongs to the technical field of enzyme-linked immunoassay. Background technique [0002] Medroxyprogesterone acetate (MPA) is a synthetic steroidal progestogen. In human pharmaceutical applications, it is used as a contraceptive in hormone replacement therapy and the treatment of hormone-dependent tumors. In veterinary applications, it can be used for animal treatment and feeding to promote growth. Due to improper use and illegal use, excessive MPA residues in animal food can inhibit human reproduction, especially pose a potential threat to children's health, and may produce various chronic and cumulative toxicity, such as "three effects ...

Claims

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Application Information

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IPC IPC(8): C07K16/18G01N33/535
Inventor 金征宇彭池方胥传来谢正军
Owner JIANGNAN UNIV
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