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Detection kit for angiotensin II and preparation method and application of detection kit

An angiotensin and anti-angiotensin technology, which is applied in the field of chemiluminescent immunoassay kits to detect the concentration of angiotensin II, can solve the problems of long time-consuming, low efficiency, and long detection time

Inactive Publication Date: 2015-05-13
SHENZHEN NEW INDS BIOMEDICAL ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, the above-mentioned radioimmunoassay has the defects of radioactive pollution, short half-life of markers, radiation damage to the operator, cumbersome operation, long time-consuming, low sensitivity, narrow detection range, and cannot be fully automated.
The traditional radioimmunoassay or enzyme-linked immunoassay method takes a long time to detect, and at the same time mainly relies on a series of cumbersome operations such as pure manual sample addition, which is inefficient and easily leads to large errors in experimental results; due to the incomplete enzymatic reaction and is susceptible to external interference factors Influence, such as temperature, time and material concentration, so the specificity of detection is low, the sensitivity is poor, and the detection range is narrow

Method used

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  • Detection kit for angiotensin II and preparation method and application of detection kit
  • Detection kit for angiotensin II and preparation method and application of detection kit
  • Detection kit for angiotensin II and preparation method and application of detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] (1) Labeling of AII antigen, the specific steps are as follows:

[0060] Preparation of dialysate (F solution): Add Na to a 5000ml container 2 CO 3 14.31g and NaHCO 3 26.46g, add purified water to dilute to 4500ml, and place the prepared F solution on a magnetic stirrer for later use.

[0061] Choose a dialysis bag with a suitable cutoff (commonly used molecular weight of 14000), measure the size enough to accommodate solution F, tie one end tightly after wetting, and test for leaks with purified water 3 times (no leakage is required).

[0062] Take 100 μg of AII antigen and adjust to 1 ml with 0.1 mol / L pH9.5 carbonic acid buffer (F solution). Put it into the dialysate, stir and dialyze at room temperature for 2 hours, add 300 μg of ABEI activated ester to the dialyzed solution, and react at 37°C for 2 hours.

[0063] The ligation product of AII antigen and ABEI was purified by G-25 gel column.

[0064] D. 2 Solution preparation: Add 200ml of 0.5M phosphate buffe...

Embodiment 2

[0081] (1) Labeling of AII antigen, the specific steps are as follows:

[0082] Preparation of dialysate (F solution): Add Na to a 5000ml container 2 CO 3 14.31g, NaHCO 3 26.46g, add purified water to dilute to 4500ml. The prepared F solution was placed on a magnetic stirrer for later use.

[0083] Choose a dialysis bag with a suitable cutoff (commonly used molecular weight of 14000), measure the size enough to accommodate solution F, tie one end tightly after wetting, and test for leaks with purified water 3 times (no leakage is required).

[0084] Take 100 μg of AII antigen and adjust to 1 ml with 0.1 mol / L pH9.5 carbonic acid buffer (F solution). Put it into the dialysate, stir and dialyze at room temperature for 2 hours, add 300 μg of ABEI activated ester to the dialyzed solution, and react at 37°C for 2 hours.

[0085] The ligation product of AII antigen and ABEI was purified by G-25 gel column.

[0086] D. 2 Solution preparation: add 200ml 0.5M phosphate buffer sa...

Embodiment 3

[0107] (1) Labeling of anti-AII antibody, the specific steps are as follows:

[0108] Preparation of dialysate (F solution): Add Na to a 5000ml beaker 2 CO 3 14.31g, NaHCO 3 26.46g, add purified water to dilute to 4500ml, and place the prepared F solution on a magnetic stirrer for later use.

[0109] Select a dialysis bag with a suitable cutoff (commonly used molecular weight of 14000), measure the size enough to accommodate solution F, tie one end tightly after wetting, and test for leaks with purified water 3 times (no leakage is required).

[0110] Take 1mg of anti-AII antibody and adjust it to 1ml with 0.1mol / L0 pH9.5 carbonic acid buffer (F solution). Put it into the dialysate, stir and dialyze at room temperature for 2 hours, add 300 μg of ABEI activated ester to the dialyzed solution, and react at 37°C for 2 hours.

[0111] The ligation product of anti-AII antibody and ABEI was purified by G-25 gel column.

[0112] Prepare D by the method in embodiment 1 2 solutio...

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Abstract

The invention relates to a chemiluminesent immunoassay kit for detecting angiotensin II and a preparation method of the chemiluminesent immunoassay kit. The kit comprises a component A and a component B, wherein the component A is angiotensin II antigen or a junctional complex of the angiotensin II antigen and a protein carrier; the component B is an angiotensin II antibody; one of the component A and the component B is used for marking a tracer marker; and the other one is enveloped by magnetic spheres. The invention further relates to a method for detecting the concentration of the angiotensin II by virtue of the kit. According to the chemiluminesent immunoassay kit, the concentration of the angiotensin II is measured by the kit provided by the invention; and high detection sensitivity and accuracy can be obtained.

Description

technical field [0001] The invention relates to a detection kit, in particular to a chemiluminescent immunoassay kit for detecting angiotensin II. The invention also relates to a preparation method of the kit and a method for using the kit to detect the concentration of angiotensin II. Background technique [0002] Angiotensin II (AII) is formed by cleavage of two amino acid residues at the C-terminus by angiotensin converting enzyme (Angiotensin0Converting Emzyme, ACE), and is an 8-peptide substance. [0003] The renin-angiotensin-aldosterone system (renin-angiotensin-aldosterone system, RAAS) is a hormone system. When massive blood loss or blood pressure drops, the kidneys secrete renin, which catalyzes the hydrolysis of angiotensinogen to produce angiotensin Ⅰ (AI), AI basically has no biological activity. AII has a high-efficiency vasoconstriction effect, thereby increasing blood pressure. AII can also stimulate the adrenal cortex to secrete aldosterone, which can pro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/74G01N33/532G01N21/76
CPCG01N33/74G01N2410/02
Inventor 饶微高莉杜凯李钦徐红罗凯李婷华袁锦云
Owner SHENZHEN NEW INDS BIOMEDICAL ENG
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