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Angiotensin I detection reagent kit as well as preparation method and application thereof

An angiotensin and kit technology, which can be used in measurement devices, analysis by chemical reaction of materials, instruments, etc., can solve the problems of radioactive contamination, low sensitivity, and narrow detection range.

Inactive Publication Date: 2015-05-20
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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  • Angiotensin I detection reagent kit as well as preparation method and application thereof
  • Angiotensin I detection reagent kit as well as preparation method and application thereof
  • Angiotensin I detection reagent kit as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] (1) Labeling of AI antigen, the specific steps are as follows:

[0061] 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.

[0062] 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).

[0063] Take 100 μg of AI antigen and adjust to 1 ml with 0.1 mol / L carbonic acid buffer (F solution) of pH 9.5. 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.

[0064] The ligation product of AI antigen and ABEI was purified by G-25 gel column.

[0065] D. 2 Solution preparation: Add 200ml of 0.5M phosphate buff...

Embodiment 2

[0082] (1) Labeling of AI antigen, the specific steps are as follows:

[0083] Dialysate (F solution) configuration: 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.

[0084] 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).

[0085] Take 100 μg of AI antigen and adjust to 1 ml with 0.1 mol / L carbonic acid buffer (F solution) of pH 9.5. 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.

[0086] The ligation product of AI antigen and ABEI was purified by G-25 gel column.

[0087] D. 2 Solution preparation: add 200ml 0.5M phosphate buffer sa...

Embodiment 3

[0109] (1) Labeling of anti-AI antibody, the specific steps are as follows:

[0110] Dialysate (F solution) configuration: 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.

[0111] 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).

[0112] Take 1 mg of anti-AI antibody and adjust it to 1 ml with 0.1 mol / L carbonic acid buffer (F solution) at pH 9.5. 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.

[0113] The ligation product of anti-AI antibody and ABEI was purified by G-25 gel column.

[0114] Prepare D by the method in embodiment 1 2 solut...

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Abstract

The invention relates to a chemiluminescence immunity detection reagent kit for detecting angiotensin I and a preparation method thereof. The reagent kit comprises a component A and a component B, wherein the component A is an angiotensin I antigen or an adapter of an angiotensin I antigen and a protein carrier, the component B is an anti-angiotensin I antibody, one of the component A and the component B is marked with a trace marker, and the other one is coated with magnetic spheres. The invention further relates to a method for detecting concentration of the angiotensin I by utilizing the reagent kit. According to the angiotensin I detection reagent kit, the concentration of the angiotensin I is determined by utilizing the reagent kit, so that the sensitivity and accuracy in detection are high.

Description

technical field [0001] The invention relates to a detection kit, in particular to a chemiluminescent immunoassay kit for detecting angiotensin I. The invention also relates to a preparation method of the kit and a method of using the kit to detect the concentration of angiotensin I. Background technique [0002] Angiotensin Ⅰ (Angiotensin Ⅰ, AI) is produced by renin acting on angiotensinogen secreted by the liver. The rate of AI production is one of the important indicators of renin activity (PRA). [0003] The renin-angiotensin-aldosterone system (renin-angiotensin-aldosterone system, RAAS) is a hormone system. When a large amount of blood loss or blood pressure drops, the kidneys secrete renin, which catalyzes the hydrolysis of angiotensinogen to produce AI. AI basically It has no biological activity, but is cleaved by angiotensin converting enzyme (Angiotensin Converting Emzyme, ACE) to form AII by cleaving two amino acid residues at the C-terminus. AII has a high-effic...

Claims

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Application Information

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IPC IPC(8): G01N33/543
CPCG01N33/54326G01N21/76G01N33/533G01N33/54353G01N2410/02
Inventor 饶微李钦徐红彭露罗凯李婷华袁锦云
Owner SHENZHEN NEW INDS BIOMEDICAL ENG
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