Adiponectin chemiluminescence immunoassay kit and its preparation method and use

A technology of chemiluminescence immunity and detection kits, which is applied in the direction of chemiluminescence/bioluminescence, anti-hormonal immunoglobulin, and analysis by making materials undergo chemical reactions, which can solve the problem of inaccurate, complicated, and complex reagent or sample additions. Problems such as poor detection accuracy

Inactive Publication Date: 2016-12-07
SHENZHEN YHLO BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The traditional detection method of adiponectin mainly adopts enzyme-linked immunosorbent assay. However, limited by the characteristics of enzyme-linked immunosorbent assay, the traditional detection method of adiponectin is in an open space during the detection process, which is easy to cause differ

Method used

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  • Adiponectin chemiluminescence immunoassay kit and its preparation method and use
  • Adiponectin chemiluminescence immunoassay kit and its preparation method and use
  • Adiponectin chemiluminescence immunoassay kit and its preparation method and use

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preparation example Construction

[0062] The preparation process of the adiponectin calibrator is as follows: the adiponectin is prepared into adiponectin solutions with a concentration of 0-200 ng / mL by using the calibrator buffer.

[0063] Specifically, the adiponectin calibration product is a solution of adiponectin with concentrations of 0 ng / mL, 0.5 ng / mL, 10 ng / mL, 20 ng / mL, 80 ng / mL and 200 ng / mL, respectively.

[0064] The adiponectin chemiluminescence immunoassay kit can use a fully automatic chemiluminescence immunoassay analyzer as a detection tool to complete the detection of adiponectin. This adiponectin chemiluminescent immunoassay kit, after experiments, has a detection sensitivity of 0.01ng / mL, which is at least 10 times more sensitive than the traditional adiponectin detection method. The detection accuracy of the kit is high.

[0065] When this adiponectin chemiluminescence immunoassay kit is used, the adiponectin calibration product is detected by a fully automatic chemiluminescence immunoa...

Embodiment 1

[0117] Example 1: Preparation of adiponectin chemiluminescent immunoassay kit

[0118] (1) Preparation of magnetic particles coated with adiponectin monoclonal antibody:

[0119] Take 50 mg of carboxylated magnetic particles (MagnaBind TM , Cat. No. 21353) suspension, magnetically separated to remove the supernatant, resuspended with 0.02M, pH 5.5 MES buffer, added 0.5mL~2mL of newly prepared 10mg / mL EDC aqueous solution to activate the carboxyl groups on the surface of the magnetic beads, and added 3mg ~5 mg of adiponectin monoclonal antibody (Novus, NB100-65810), suspended at room temperature for 2h~10h, magnetically separated, removed the supernatant, and resuspended with 0.1M Tris buffer at pH 8.0 containing 2% BSA to 1mg / mL to obtain adiponectin monoclonal antibody-coated magnetic particles, and store in 5mL bottles at 4°C for future use.

[0120] (2) Preparation of adiponectin monoclonal antibody labeled with acridinium ester:

[0121] Take 500 μL 1.0 mg / mL adiponecti...

Embodiment 2

[0124] Example 2: Chemiluminescent immunoassay method for adiponectin acridinium ester

[0125] The present invention uses a full-automatic chemiluminescence immunoassay analyzer and the adiponectin chemiluminescence immunoassay kit prepared in Example 1 as detection tools. Magnetic particles coated with adiponectin monoclonal antibody and 50 μL of acridinium ester-labeled adiponectin monoclonal antibody were reacted for 10 minutes and then magnetically separated. The instrument sent the reaction mixture into a dark room, and sequentially added luminescent substrate A solution ( Contains 0.1M HNO 3 and 0.5%H 2 o 2 ) and B solution (containing 0.25M NaOH) for luminescence reaction, and finally record the luminescence intensity, and calculate the adiponectin content of the tested sample from the standard curve.

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Abstract

The invention discloses an adiponectin chemiluminescence immunoassay kit and its preparation method and use. The adiponectin chemiluminescence immunoassay kit comprises an adiponectin monoclonal antibody-coated solid phase carrier and a chemiluminescent label-labeled adiponectin monoclonal antibody. The adiponectin chemiluminescence immunoassay kit utilizes a full-automatic chemical luminescence immunoassay instrument as a detection tool to finish adiponectin detection. An experiment proves that the adiponectin chemiluminescence immunoassay kit has detection sensitivity of 0.01ng/mL. Compared with the traditional adiponectin detection method, the adiponectin chemiluminescence immunoassay kit improves sensitivity by at least 10 times and has a high detection precision.

Description

technical field [0001] The invention relates to the field of in vitro detection, in particular to an adiponectin chemiluminescent immunoassay kit and its preparation method and application. Background technique [0002] Adipose tissue is mainly composed of a large number of adipocytes aggregated into agglomerates. Adiponectin (ADPN) is an endogenous bioactive polypeptide or protein secreted by adipocytes. It appears in the human body at a concentration of 3-30ug / ml. in circulating plasma. Adiponectin is also known as Acrp30, apM1, AdipoQ, GBP28. Initially, adiponectin was found in human subcutaneous adipose tissue, plasma and adipocytes of murine animals. Adiponectin in the human body consists of 244 amino acids with a molecular weight of 30KD. It consists of an amino-terminal secretion signal sequence (aa 1-18), a specific sequence (aa19-41), a group of 22 amino acid repeat collagen sequences (aa 42-107), and a globular sequence (aa108-244) . Among them, the globular re...

Claims

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Application Information

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IPC IPC(8): G01N33/74G01N33/532G01N21/76
CPCG01N21/76G01N33/532G01N33/74G01N33/582C07K16/26G01N33/54306G01N33/54326
Inventor 夏福臻钱纯亘王刚祝亮何雨禧
Owner SHENZHEN YHLO BIOTECH
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