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Renin chemiluminescence detection kit

A technology of chemiluminescence detection and kits, which is applied in the direction of chemiluminescence/bioluminescence, analysis by causing chemical reactions of materials, and measurement devices, which can solve the problems of large errors, low degree of automation, and high cost of reagents

Active Publication Date: 2018-08-14
SICHUAN MACCURA BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The plate chemiluminescence method is similar to the enzyme-linked immunosorbent method, with a low degree of automation and mostly manual operations. The detection results have large errors due to human factors and low sensitivity; the magnetic particle chemiluminescence method uses fully automatic instruments for detection, and the degree of automation Higher and more stable results, but the kits currently used in clinical practice generally have problems of low sensitivity and insufficient low-end resolution, and multi-point calibrator is often used for calibration, which increases the cost of reagent testing and the burden on patients
[0005] In China, renin detection is mainly based on chemiluminescence immunoassay in clinical practice. However, due to the fact that there are still relatively few domestic renin detection kits with independent intellectual property rights, and the sensitivity is low and the cost of reagents is high. It has been carried out in tertiary hospitals, but less in hospitals below the tertiary level. Therefore, a detection technology with high sensitivity, good accuracy and low detection cost for renin detection needs to be developed.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1: Preparation of avidinized magnetic particles (reagent 1) coated with renin monoclonal antibody

[0020] 1. Preparation of Biotinylated Renin Monoclonal Antibody

[0021] Take 1.0 mg of renin monoclonal antibody, dilute to 0.5 mg / mL with phosphate buffer, then add 13.3 μL of biotin ester with a concentration of 2.5 mM / L, react at room temperature for 30 min, and then add 20 μL of Tris solution with a concentration of 1M , reacted at room temperature for 10 minutes, and finally dialyzed with 20 mM phosphate buffer solution with pH 7.4 for 16-24 hours, and obtained the biotinylated renin monoclonal antibody by taking the dialyzed liquid.

[0022] 2. Biotinylated antibody coated avidin magnetic particles

[0023] Take 50mg of the avidinized magnetic particle suspension, magnetically separate to remove the supernatant, and use ligation buffer (20mMPBS, 10g / LBSA, 20g / L mannitol, 10mL / L glycerol, 0.1mL / L TritonX-100, 1mL / L proclin-300, 20mg / L 4-aminoantipyrine) wa...

Embodiment 2

[0025] Example 2: Preparation of acridinium ester-labeled renin monoclonal antibody (reagent 2)

[0026] Take 1.0mg of renin monoclonal antibody, dilute it to 2.0mg / mL with 20mM phosphate buffer, then add 20μL of acridinium ester with a concentration of 10mM / L, mix and react for 30min in the dark at room temperature, and then add 100μL of a concentration of 10mg / L mL of lysine solution, mixed and reacted for 10 min in the dark at room temperature, and then desalted and purified by centrifugation with a desalting column. Collect the liquid in the centrifuge tube to obtain the acridinium ester-labeled renin monoclonal antibody. Then dilute the acridinium ester-labeled renin monoclonal antibody obtained above to a concentration of 4.0 μg / mL with a marker diluent to obtain reagent 2. The marker diluent is a 20 mM PBS solution with a pH of 7.4, containing 10 g / L Casein, 10g / L trehalose, 10mL / L glycerin, 0.1mL / L TritonX-100, 1mL / L proclin-300 and 20mg / L 4-aminoantipyrine. In addit...

Embodiment 3

[0027] Embodiment three: preparation of renin calibrator

[0028] Prepare the renin antigen with the calibrator diluent to a concentration of 10 μIU / mL and 100 μIU / mL, divide into 1.0 mL / bottle and freeze-dry. The renin calibrator can be traced to the WHO international standard (WHO International Standard RENIN, Human, NIBSC code: 68 / 356). Calibrator dilution 20mM pH7.4 is a PBS solution containing 10g / L casein, 50g / L bovine serum albumin, 20g / L trehalose, 10mL / L glycerol, 0.1mL / L TritonX-100, proclin- 300 1mL / L and 20mg / L 4-aminoantipyrine.

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Abstract

The invention discloses a renin chemiluminescence detection kit which comprises a reagent 1 and a reagent 2. The reagent 1 is prepared from a buffer solution, magnetic micro particles, a biotin labelled anti-renin monoclonal antibody and polyhydroxy alcohol; the reagent 2 is prepared from a buffer solution, an acridinium ester labelled anti-renin monoclonal antibody and polyhydroxy saccharide. Therenin chemiluminescence detection kit provided by the invention is high in sensitivity, good in accuracy and capable of reducing the detection cost.

Description

technical field [0001] The invention relates to the field of in vitro diagnostic reagents, in particular to a renin chemiluminescence detection kit. Background technique [0002] Renin (English: Renin), also known as angiotensinogenase, is a proteolytic enzyme released by the juxtaglomerular cells of the juxtaglomerular apparatus (also known as the juxtaglomerular complex). component of the element system. Renin enters the blood through the renal vein, and can catalyze the conversion of angiotensinogen secreted by the liver into the plasma into angiotensin Ⅰ, and increase the production of aldosterone. Increased renal tubular reabsorption of NaCl and water converts enzymes in the blood and lung tissue to degrade angiotensin I to angiotensin II, which can be hydrolyzed by aminopeptidases to angiotensin III. All three angiotensins have biological activity, among which angiotensin II and III have stronger biological activity, and the concentration of the latter in blood is lo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/76
CPCG01N21/763
Inventor 赵浩瀚郭鹏黄燕玲田君喜龙腾镶
Owner SICHUAN MACCURA BIOTECH CO LTD
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