Serum amyloid protein A detection kit and preparation method thereof

A detection kit and serum starch technology, which is applied in the field of medical detection, can solve the problems of large amount of antibody, loss of binding force of antibody, and susceptibility to interference, etc., so as to improve the efficiency of antibody coating, accurate binding position, and improve anti-interference ability Effect

Pending Publication Date: 2020-03-24
SICHUAN XINCHENG BIOLOGICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

As stated in the patent CN105372434A: (1) the sensitized latex particles obtained by the physical adsorption method, the antibody is easy to fall off from the latex particles, and the latex obtained by the physical adsorption method is easily disturbed, resulting in false positive or false detection results. Elevated Negative
(2) The chemical coupling method is mostly random coupling method, because the antibody is randomly coupled to different parts of the latex, which will lead to the loss of binding force of the antibody
The disadvantages of this method are: the preparation process is more complicated, the centrifugation time is long, and the amount of antibody is large, which increases the production cost

Method used

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  • Serum amyloid protein A detection kit and preparation method thereof
  • Serum amyloid protein A detection kit and preparation method thereof
  • Serum amyloid protein A detection kit and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] The serum amyloid A detection kit of the present invention comprises the following reagents:

[0040] Reagent 1: 4-hydroxyethylpiperazineethanesulfonic acid 50mMol / L, Tween-20 5ml / L~15ml / L, Brij-35 5g / L-10g / L, sodium chloride 10g / L, polyethylene glycol 8000 10g / L, sodium azide 1g / L, pH value 7.0;

[0041] Reagent 2: 4-hydroxyethylpiperazineethanesulfonic acid 50mM, latex particles coated with mouse anti-human SAA monoclonal antibody, mouse anti-human SAA monoclonal antibody, sodium borohydride 0.05g / L, casein 2g / L, Bovine serum albumin 5g / L, glucose 80g / L, sodium azide 1g / L.

[0042] The latex particles coated with the mouse anti-human SAA monoclonal antibody are polystyrene microspheres modified by aldehyde groups.

Embodiment 2

[0044] Serum amyloid A detection kit of the present invention is prepared through the following steps:

[0045] Reagent 1:

[0046] (1) Add 4-hydroxyethylpiperazineethanesulfonic acid, Tween-20, Brij-35, sodium chloride, polyethylene glycol 8000 and sodium azide successively in the reactor while stirring, stir until completely dissolved, Make the concentration of each component reach as follows: 4-hydroxyethylpiperazineethanesulfonic acid 50mM, Tween-20 5ml / L~15ml / L, Brij-35 5g / L-10g / L, sodium chloride 10g / L, Polyethylene glycol 800010g / L, sodium azide 1g / L, pH value 7.0;

[0047] Reagent 2:

[0048] (1) prepare 4-hydroxyethylpiperazine ethanesulfonic acid solution;

[0049] (2) Dilute the latex particles coated with the mouse anti-human SAA monoclonal antibody to 1% with the above (1) solution, and then preheat the solution to 34-37°C;

[0050] (3) Add all the mouse anti-human SAA monoclonal antibody dropwise to the solution of (2), mix well, and react for 30 minutes;

[0...

Embodiment 3

[0057] Serum amyloid A detection kit, including the following reagents:

[0058] Reagent 1: 4-hydroxyethylpiperazineethanesulfonic acid 50mMol / L, Tween-20 5ml / L~15ml / L, Brij-35 5g / L-10g / L, sodium chloride 10g / L, polyethylene glycol 8000 10g / L, sodium azide 1g / L, pH value 7.0;

[0059] Reagent 2: 4-hydroxyethylpiperazineethanesulfonic acid 50mM, latex particles coated with mouse anti-human SAA monoclonal antibody, mouse anti-human SAA monoclonal antibody, EDAC 0.1g / L, casein 2g / L, bovine serum Albumin 5g / L, glucose 80g / L, sodium azide 1g / L.

[0060] The difference between this example and Example 1 is that the latex particles coated with the mouse anti-human SAA monoclonal antibody use conventional microspheres, the reagent 2 does not contain sodium borohydride, and the activator uses EDAC.

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Abstract

The invention discloses a serum amyloid protein A detection kit which comprises a reagent 1 and a reagent 2, and the reagent 2 comprises latex particles coated with a mouse anti-human SAA monoclonal antibody, the mouse anti-human SAA monoclonal antibody, sodium borohydride, bovine serum albumin and bovine serum albumin. According to the kit, on the basis of a chemical coupling method, aldehyde microspheres are adopted, so that the binding force of an antibody and latex is improved, and the antibody coating efficiency is further improved.

Description

technical field [0001] The invention relates to the field of medical detection, in particular to a serum amyloid A detection kit and a preparation method thereof. Background technique [0002] Serum amyloid A protein (serumamyloid A protein, SAA) is an acute phase reaction protein produced by hepatocytes and then secreted into serum. Lipoprotein (HDL) binding. 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, when the body's antigens are cleared, it will quickly drop to normal levels. Compared with CRP, which is the most widely used clinically at present, there is one most important difference between SAA: the increase of SAA is seen in virus, mycoplasma, and bacterial infection, and its sensitivity is higher than that of CRP; the increase of CRP is se...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N33/577G01N33/68
CPCG01N33/54313G01N33/577G01N33/68
Inventor 林源万雅心林婷婷蒲建文
Owner SICHUAN XINCHENG BIOLOGICAL CO LTD
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