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Kit for quantitative detection on serum amyloid protein A and preparation method

A serum starch, quantitative detection technology, applied in the field of biomedicine, can solve the problems of poor specificity, low detection sensitivity, narrow detection range, etc., and achieve the effect of high accuracy, high sensitivity and high sensitivity

Inactive Publication Date: 2018-06-15
浙江艾明德生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] This solution provides a chemiluminescence detection reagent and preparation method for the quantitative detection of serum amyloid A, which solves the shortcomings of low detection sensitivity, poor specificity, and narrow detection range in the prior art.

Method used

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  • Kit for quantitative detection on serum amyloid protein A and preparation method
  • Kit for quantitative detection on serum amyloid protein A and preparation method
  • Kit for quantitative detection on serum amyloid protein A and preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1 Preparation of Antibodies to Serum Amyloid A

[0041] SAA monoclonal antibody, dialyzed overnight at 4°C with 0.05M pH 7.5 phosphate buffered saline (PBS).

Embodiment 2

[0042] Example 2 Preparation of the serum amyloid A quantitative assay kit of the present invention

[0043] 1. Preparation of horseradish peroxidase antibody

[0044] 1. Horseradish peroxidase-conjugated monoclonal antibody diluent:

[0045] Phosphate buffered saline (PBS): 0.2% 0.02M bovine serum albumin (BSA) at pH 7.5 by weight.

[0046] 2. The concentration of horseradish peroxidase-labeled monoclonal antibody is selected:

[0047] The working concentration of horseradish peroxidase-labeled monoclonal antibody was 0.2 μg / mL.

[0048] 2. Preparation of SAA calibrator

[0049]Prepare with pure SAA, dilute with diluent, pack into 0.5mL / bottle, concentration is 0, 5, 20, 60, 200, 600ng / mL, 6 bottles in total, store at 2-8°C.

[0050] 3. Preparation of antibody labeled with horseradish peroxidase:

[0051] (1) Weigh 5mg of horseradish peroxidase (HRP), dissolve it in 1ml of distilled water to form a HRP solution with a concentration of 5mg / ml;

[0052] (2) Add 0.2ml of 0...

Embodiment 3

[0078] Embodiment 3 Coated microplate technology:

[0079] Chemiluminescence refers to the emission of light accompanying the chemical reaction process. During the chemical reaction, certain substances (luminescent agents) absorb the chemical energy generated during the reaction, causing the electrons in the product molecules of the reaction or the intermediate state molecules of the reaction to jump to the excited state. When the electrons return from the excited state to the ground state , energy is released in the form of photons.

[0080] In the chemiluminescence process, the coating concentration in the coating process is different, and the ion concentration and pH value of the buffer have a great influence on the adsorption capacity of the coated microplate, the adsorption effect and the consistency between the wells, thus Affect the precision, repeatability and accuracy of the entire kit.

[0081] 1. Serum amyloid A coating concentration screening

[0082] The presen...

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Abstract

The invention provides a kit for quantitative detection on a serum amyloid protein A. The kit comprises a horse radish peroxidase marked monoclonal antibody of a serum amyloid protein A, a carrier wrapped by the monoclonal antibody of the serum amyloid protein A, a serum amyloid protein A calibration product, and a chemiluminiscence substrate. A preparation method of the kit comprises the following steps: marking the monoclonal antibody of the serum amyloid protein A by using horse radish peroxidase; wrapping the carrier by the monoclonal antibody of the serum amyloid protein A; preparing theserum amyloid protein A calibration product by using a pure serum amyloid protein A product; subpackaging the serum amyloid protein A calibration product, the horse radish peroxidase marked monoclonalantibody of the serum amyloid protein A, and the chemiluminiscence substrate under the action of HRP (Horse Radish Peroxidase); preparing a finished product through assembling. The kit provided by the invention is high in sensitivity, good in specificity, high in quantitative detection result accuracy, low in use cost and relatively easy to popularize and apply.

Description

[0001] The invention relates to the field of biomedicine, in particular to a kit and a preparation method for quantitatively detecting serum amyloid A. Background technique [0002] Serum amyloid A (serum amyloid A, SAA) is an acute time-limited response protein, which belongs to the heterogeneous protein in the apolipoprotein family, with a relative molecular weight of about 12,000. In an acute time-limited reaction, stimulated by IL-1, IL-6 and TNF, SAA is synthesized in the liver by activated macrophages and fibroblasts, which can increase to 100-1000 times the initial concentration, but the half-life is extremely short. Short, only about 50 minutes. Serum amyloid A is associated with high-density lipoprotein (HDL), which regulates HDL metabolism during inflammation. A particularly important property of serum amyloid A is that its degradation products can be deposited in different organs as amyloid A (AA) fibrils, which is a serious complication in chronic inflammatory dis...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/76G01N33/68G01N33/577
CPCG01N21/76G01N33/577G01N33/6893G01N2800/7095
Inventor 胡国富朱炎
Owner 浙江艾明德生物科技有限公司
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