Chemiluminescence immunoassay assay determination kit for detecting human growth hormone and preparation method thereof

A chemiluminescence immunoassay and human growth hormone technology, applied in the field of immunoassay, can solve the problems of poor repeatability, long detection time, low sensitivity, etc.

Inactive Publication Date: 2018-11-09
DIRUI MEDICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] The purpose of the present invention is to provide a chemiluminescence immunoassay assay kit for detecting human growth hormone and its preparation method in order to solve the problems of long detection time, low sensitivity and poor repeatability of the existing methods for detecting growth hormone

Method used

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  • Chemiluminescence immunoassay assay determination kit for detecting human growth hormone and preparation method thereof
  • Chemiluminescence immunoassay assay determination kit for detecting human growth hormone and preparation method thereof
  • Chemiluminescence immunoassay assay determination kit for detecting human growth hormone and preparation method thereof

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preparation example Construction

[0041] The present invention also includes a preparation method for detecting human growth hormone chemiluminescence immunoassay assay kit, the specific process is as follows: figure 1 shown, including:

[0042] Step 1: Preparation of magnetic bead suspension coated with streptavidin

[0043] After mixing the streptavidin magnetic particle solution and TBST solution, place it on a magnetic separator until the supernatant is free of turbidity, discard the supernatant, keep the magnetic particles, and prepare a solid phase reagent in the buffer after washing R1; the concentration of the streptavidin magnetic particle solution is preferably 100mg / ml; the volume ratio of the streptavidin magnetic particle solution and TBST solution is preferably 0.5:10; the buffer is 50mM MES, 0.05% temperature, 0.05% Proclin300, pH6.0; the concentration of the magnetic beads coated with streptavidin in the solid-phase reagent is preferably 0.05%-0.1%;

[0044] Step 2: Preparation of acridinium ...

Embodiment 1

[0053] Embodiment 1 prepares GH chemiluminescent detection kit of the present invention

[0054] 1. Preparation of calibrator

[0055] Dilute GH into a calibrator with 100mM PBS buffer containing 20% ​​calf serum, aliquot into 0.03ng / ml, 0.1ng / ml, 0.5ng / ml, 1ng / ml, 2ng / ml, 5ng / ml, 10ng / ml, 25ng / ml, 50ng / ml.

[0056] 2. Preparation of R1 solution coated with streptavidin magnetic beads

[0057] Take 0.5 ml (50 mg) of streptavidin magnetic particle solution with a concentration of 100 mg / ml, add 10 ml of TBST solution and mix well for 10 minutes, then place it on a magnetic separator until the supernatant is clear of turbidity, discard the supernatant , to retain the magnetic particles. After repeated washing for 3 times, a solid-phase reagent with a magnetic bead concentration of 0.05% was formulated in a buffer solution of 50mM MES, 0.05% Tween, 0.05% Proclin300, pH 6.0, and stored at 2-8°C.

[0058] 3. Preparation of acridinium ester-labeled GH monoclonal antibody R2 sol...

Embodiment 2

[0062] Embodiment 2 Preparation of GH chemiluminescence detection kit of the present invention

[0063] 1. Preparation of calibrator

[0064] Dilute GH into a calibrator with 100mM PBS buffer containing 20% ​​calf serum, aliquot into 0.03ng / ml, 0.1ng / ml, 0.5ng / ml, 1ng / ml, 2ng / ml, 5ng / ml, 10ng / ml, 25ng / ml, 50ng / ml.

[0065] 2. Preparation of R1 solution coated with streptavidin magnetic beads

[0066] Take 0.5 ml (50 mg) of streptavidin magnetic particle solution with a concentration of 100 mg / ml, add 10 ml of TBST solution and mix well for 10 minutes, then place it on a magnetic separator until the supernatant is clear of turbidity, discard the supernatant , to retain the magnetic particles. After repeated washing for 3 times, a solid-phase reagent with a magnetic bead concentration of 0.03% was formulated in a buffer solution of 50mM MES, 0.05% Tween, 0.05% Proclin300, pH 6.0, and stored at 2-8°C.

[0067] 3. Preparation of acridinium ester-labeled GH monoclonal antibody...

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Abstract

The invention provides a chemiluminescence immunoassay assay determination kit for detecting human growth hormone and a preparation method thereof, belonging to the technical field of immunoassay. Thekit contains a magnetic bead suspension liquid coated with streptavidin, acridinium ester labeled GH antibody and biotin labeled GH antibody. The invention also provides a preparation method of the chemiluminescence immunoassay assay determination kit for detecting human growth hormone. For the kit, a sandwich method principle is adopted for quantitatively determining GH in serum or plasma, a biotin-GH antibody compound is added to streptavidin-magnetic bead suspension liquid, a magnetic bead-avidin-biotin-GH compound is formed by the affiliation reaction of biotin and streptavidin, and an antigen-antibody compound is formed by antigen-antibody reaction to form magnetic bead-biotin-GH antibody-GH-GH antibody-acridinium ester. The detection method has the advantages of high sensitivity, strong specificity, good repeatability, wide detection scope and relatively low cost, and can be widely applied to clinical detection.

Description

technical field [0001] The invention belongs to the technical field of immune analysis, and relates to a chemiluminescence immunoassay assay kit for detecting human growth hormone and a preparation method thereof. Background technique [0002] Human Growth Hormone is a single peptide chain protein hormone secreted by eosinophilic cells in the anterior pituitary gland. It consists of 191 amino acids and has a wide range of physiological functions. Human Growth Hormone mainly stimulates the liver to secrete insulin-like growth factors. (IGF-I), acts on bone and cartilage, causing height growth, and at the same time can promote human protein synthesis and fat decomposition, and regulate water and salt balance in the body. [0003] 1. Clinical manifestations of auxin abnormalities [0004] Growth hormone abnormalities are mainly divided into: 1) growth hormone excess; 2) growth hormone deficiency. Gigantism and acromegaly are commonly seen with growth hormone excess. Gigantis...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/74
Inventor 翟涛刘杨何浩会高威孙成艳
Owner DIRUI MEDICAL TECH CO LTD
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