Method, reagent strip and kit for labeling typhoid salmonella recombinant antigen by colloidal gold

A technology of Salmonella typhi and recombinant antigen, applied in the field of gold immunity, can solve the problem of not being able to significantly improve the color intensity and sensitivity of the product

Active Publication Date: 2021-04-09
山东康华生物医疗科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] The characteristics of immune gold are mainly affected by gold particles, the dosage of recombinant Salmonella typhi antigen, and the pH value. The optimal dosage of recombinant Salmonella typhi antigen is generally the maximum amount of recombinant antigen labeled Salmonella typhi completely adsorbed on colloidal gold, and the optimum pH is The value is generally close to or slightly higher than the isoelectric point of the Salmonella typhi recombinant antigen to be labeled, but the optimal labeling state can only ensure the stability of immunogold, and cannot significantly improve the color intensity and sensitivity of the product in immunochromatography

Method used

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  • Method, reagent strip and kit for labeling typhoid salmonella recombinant antigen by colloidal gold

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Embodiment 1: A kind of method for colloidal gold labeling Salmonella typhi recombinant antigen, labeling method comprises the following steps:

[0041] (1) Liquid preparation

[0042] Chlorauric acid solution: Dissolve 10 g of chloroauric acid in ultrapure water, and after it is completely dissolved, dilute to 1000 mL, and store in the dark at 4°C for later use.

[0043] Trisodium citrate solution: Weigh 1 g of trisodium citrate, dissolve it in 100 mL of ultrapure water at room temperature, filter through a 0.22 μm membrane filter, and store at 4°C for later use.

[0044] Potassium stannate solution: Weigh 2 g of potassium stannate, dissolve it in 100 mL of ultrapure water at room temperature, filter it with a 0.22 μm filter membrane, and store it at 4°C for later use.

[0045] BSA solution: Weigh 10g of BSA, dissolve in 100mL of ultrapure water at room temperature, store at 4°C, and prepare immediately after use.

[0046] PEG20000 solution: Weigh 10g of PEG20000, di...

Embodiment 2

[0052] Embodiment 2: A kind of method for colloidal gold labeling Salmonella typhi recombinant antigen, labeling method comprises the following steps:

[0053] (1) Liquid preparation

[0054] Chlorauric acid solution: Dissolve 10 g of chloroauric acid in ultrapure water, and after it is completely dissolved, dilute to 1000 mL, and store in the dark at 4°C for later use.

[0055]Trisodium citrate solution: Weigh 1 g of trisodium citrate, dissolve it in 100 mL of ultrapure water at room temperature, filter through a 0.22 μm membrane filter, and store at 4°C for later use.

[0056] Potassium stannate solution: Weigh 2 g of potassium stannate, dissolve it in 100 mL of ultrapure water at room temperature, filter it with a 0.22 μm filter membrane, and store it at 4°C for later use.

[0057] BSA solution: Weigh 10g of BSA, dissolve in 100mL of ultrapure water at room temperature, store at 4°C, and prepare immediately after use.

[0058] PEG20000 solution: Weigh 10g of PEG20000, dis...

Embodiment 3

[0064] Embodiment 3: a kind of method for colloidal gold labeling Salmonella typhi recombinant antigen, labeling method comprises the following steps:

[0065] (1) Liquid preparation

[0066] Chlorauric acid solution: Dissolve 10 g of chloroauric acid in ultrapure water, and after it is completely dissolved, dilute to 1000 mL, and store in the dark at 4°C for later use.

[0067] Trisodium citrate solution: Weigh 1 g of trisodium citrate, dissolve it in 100 mL of ultrapure water at room temperature, filter through a 0.22 μm membrane filter, and store at 4°C for later use.

[0068] Potassium stannate solution: Weigh 2 g of potassium stannate, dissolve it in 100 mL of ultrapure water at room temperature, filter it with a 0.22 μm filter membrane, and store it at 4°C for later use.

[0069] BSA solution: Weigh 10g of BSA, dissolve in 100mL of ultrapure water at room temperature, store at 4°C, and prepare immediately after use.

[0070] PEG20000 solution: Weigh 10g of PEG20000, di...

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Abstract

The invention relates to a method for labeling a typhoid salmonella recombinant antigen by colloidal gold. The method comprises the following steps: (1) preparing a colloidal gold solution; (2) adjusting the pH value of the prepared colloidal gold solution to be lower than the optimal pH value by 1.5-2.0; (3) adding the labelled typhoid salmonella recombinant antigen at one time, wherein the adding amount is 70%-80% lower than the optimal use amount of the labelled typhoid salmonella recombinant antigen; and (4) adding a sealing agent and a stabilizing agent, and finishing labeling. The safety range is broken, the color development intensity and sensitivity of the product can be obviously improved by placing immune gold under a relatively unstable and controllable condition, the 'relatively unstable and controllable condition' means that the pH value labelled by colloidal gold is slightly lower than the isoelectric point labelled by typhoid salmonella recombinant antigen, and the use amount of the labelled typhoid salmonella recombinant antigen is unsaturated; and the sealing agent and the stabilizer are added at the same time, so that the color development intensity of the product is obviously deepened, and the sensitivity is obviously improved.

Description

technical field [0001] The invention belongs to the technical field of gold immunization, and in particular relates to a method for colloidal gold labeling Salmonella typhi recombinant antigen, a reagent strip and a kit. Background technique [0002] Gold immune technology is an immunolabeling technology using colloidal gold as a marker. Colloidal gold is a negatively charged hydrophobic colloid solution formed by chloroauric acid under the action of a reducing agent. The solution becomes a stable colloidal state due to electrostatic interaction , which is colloidal gold, the surface of colloidal gold is negatively charged, and the positively charged groups of proteins are coated on the surface of colloidal gold particles due to electrostatic adsorption. This process is colloidal gold labeling, also known as gold immunity. [0003] The characteristics of immune gold are mainly affected by gold particles, the dosage of recombinant Salmonella typhi antigen, and the pH value. T...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/532G01N33/543G01N33/558G01N33/569
CPCG01N33/532G01N33/54313G01N33/558G01N33/56916G01N2333/255
Inventor 杨明霞杨盛林戴晓华杨帆梁莉甜范文翠
Owner 山东康华生物医疗科技股份有限公司
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