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Animal progesterone chemiluminescence detection kit

A chemiluminescence detection and kit technology, applied in the field of immunoassay, can solve the problems of long detection period of radioimmunoassay, cannot be used to detect progesterone, and high technical requirements for operation, and achieves stable enzyme conjugates, long platform period, Simple effect of adding samples

Inactive Publication Date: 2019-03-08
CHENGDU POLYTECH BIOLOGICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Chromatography, as a standard method for detecting progesterone in serum, has incomparable advantages in terms of accuracy, selectivity, repeatability, and sensitivity. Pre-processing, etc.
[0004] 1. The above progesterone detection method can only be applied to drug residues or human diagnosis, but not to the detection of progesterone in animal serum or plasma
[0005] 2. The radioimmunoassay method has a long detection cycle, a short validity period, and is prone to radioactive contamination
Enzyme-linked immunoassay has a low degree of automation, and the reaction time is long, and the detection efficiency is low

Method used

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  • Animal progesterone chemiluminescence detection kit
  • Animal progesterone chemiluminescence detection kit
  • Animal progesterone chemiluminescence detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Preparation of capillary tubes coated with progesterone monoclonal antibody:

[0056] Modified progesterone monoclonal antibody, with a sulfhydryl group attached to the amino group of the protein:

[0057] The concentration of the progesterone monoclonal antibody to be coated is 2mg / mL, and the amount of 2-IT used is twice the molar amount of the progesterone monoclonal antibody. The two are mixed in 0.01M PBS (pH 8) and reacted at room temperature for 0.5h. Unreacted 2-IT was removed through a desalting column, and the buffer was replaced with 0.01M PBS (pH 6.5).

[0058] Connect the capillary to the amino group through a modifier, and use SMCC or its derivatives to activate the amino group on the modified capillary:

[0059] SMCC was first dissolved in dimethyl sulfoxide or N,N-dimethylformamide or other organic solvents, then dissolved in 0.01M PBS (pH 7), added to the modified capillary and submerged, the concentration of SMCC was 0.1mg / mL. React at room tempera...

Embodiment 2

[0075] The difference from Example 1 is that the activated SMCC amino group on the progesterone monoclonal antibody is used to couple with the capillary-modified sulfhydryl group to obtain a capillary coated with the progesterone monoclonal antibody.

Embodiment 3

[0077] This example is basically the same as Example 1, except that the carboxyl group of the progesterone monoclonal antibody is coupled to the amino group on the ammoniated glass capillary via carbodiimide to obtain a capillary coated with the progesterone monoclonal antibody. The auxiliary reagent is N-hydroxysuccinimide, which improves the stability of the intermediate reaction product.

[0078] Activation of the carboxyl group on the progesterone monoclonal antibody with carbodiimide and N-hydroxysuccinimide:

[0079] The concentration of progesterone monoclonal antibody to be activated was 1 mg / mL, the concentration of carbodiimide was 1 mM, the concentration of N-hydroxysuccinimide was 2 mM, and the buffer was 0.01-0.1M MES (pH 5). React at room temperature for 15 min. Unreacted carbodiimide and N-hydroxysuccinimide were removed through a desalting column, and the buffer was replaced with 0.01M PBS (pH 7.0).

[0080] The activated progesterone monoclonal antibody was ...

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Abstract

The invention discloses an animal progesterone chemiluminescence detection kit which comprises a capillary wrapping a progesterone monoclonal antibody, an enzyme conjugate, an enhanced chemiluminescent and cleaning solution. The kit is high in detection efficiency, high in sensitivity and automation degree, high in specificity, simple and rapid to operate, wide in detection range, free from pollution and long in expiration date and can effectively improve current deficiency of products on the market. The kit is used for detecting the concentration of progesterone in animal serum or plasma.

Description

technical field [0001] The invention belongs to the technical field of immune analysis, in particular to a chemiluminescence detection kit for animal progesterone. Background technique [0002] Progesterone (P4) is a reproductive hormone that participates in the regulation of mammalian estrus, germ cell growth, sexual behavior, fertilization, embryo implantation, fetal development, etc., and plays an important role in the normal reproduction of animals. Progesterone is often used as one of the indicators for estrus and pregnancy status judgment. With the rapid development of the pet market, animal breeding and animal medical care need to detect the estrus state of animals, determine the ovulation time of animals, improve animal reproductive ability, and reduce the empty pregnancy rate. [0003] At present, the commonly used progesterone detection methods include radioimmunoassay, ELISA technology, and colloidal gold chromatography diagnostic technology. Chromatography, as ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577
CPCG01N33/577
Inventor 肖红冉鹏
Owner CHENGDU POLYTECH BIOLOGICAL TECH CO LTD
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