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Immunochromatographic test paper for detecting hepatitis B surface antigens and preparation method thereof

A technology of hepatitis B surface antigen and immunochromatographic test paper, which is applied in biological testing, coatings, measuring devices, etc., can solve the problems of limiting superparamagnetic composite particles, long color development time, cumbersome and complicated problems, etc.

Inactive Publication Date: 2014-02-05
昆明云大生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these preparation and modification methods are often cumbersome and complicated, and the obtained superparamagnetic composite particles cannot meet the requirements of LFIAs in terms of size, biocompatibility, saturation magnetic strength, external magnetic field response speed, stability, and labeling efficiency. The size of the beads is mostly larger than 300nm. Due to the large size of the magnetic beads, the swimming time on the test paper is slow and the color development time is long; and the particles that are too small cannot provide sufficient magnetic resonance signals; in addition, there are biocompatibility. Stability, easy aggregation of magnetic beads and other issues; these shortcomings limit the application of superparamagnetic composite particles in LFIAs

Method used

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  • Immunochromatographic test paper for detecting hepatitis B surface antigens and preparation method thereof
  • Immunochromatographic test paper for detecting hepatitis B surface antigens and preparation method thereof
  • Immunochromatographic test paper for detecting hepatitis B surface antigens and preparation method thereof

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Experimental program
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Effect test

Embodiment 1

[0053] (1) Preparation of anti-HBsAg antibody labeled with superparamagnetic composite particles

[0054] Superparamagnetic Fe with a particle size of 100nm, a particle size deviation of 15%, a magnetic saturation intensity of 40emu / g, a corresponding external magnetic field response speed of 20 seconds, and a surface carboxyl content of 80μmol / g was used. 3 o 4 Nanoparticles.

[0055] The specific method is: take 2.5mg of the above-mentioned superparamagnetic Fe 3 o 4 Nanoparticles were washed with MES buffer solution with a concentration of 0.1 mol and a pH of 4.7, and after separation and enrichment with a 0.4T magnetic rack, 1 ml of the above-mentioned MES buffer solution was added to resuspend, and then 0.96 mg of EDC and 2.17 mg of NHS were added. Mix well, react at a reaction temperature of 37° C. for 0.5 hour, and then wash with a borax buffer solution with a concentration of 50 mmol and a pH of 8.5 to obtain 2.5 mg of activated magnetic particles.

[0056] Take 0....

Embodiment 2

[0062] (1) Preparation of anti-HBsAg antibody labeled with superparamagnetic composite particles

[0063] Superparamagnetic Fe with a particle size of 60nm, a magnetic saturation intensity of 80emu / g, a corresponding external magnetic field response speed of 100 seconds, and a surface carboxyl content of 50μmol / g was used. 3 o 4 Nanoparticles.

[0064] The specific method is: take 2.5mg of the above-mentioned superparamagnetic Fe 3 o 4 Nanoparticles were washed with MES buffer solution with a concentration of 0.1 mol and a pH of 4.7, and separated and enriched with a 0.4T magnetic rack, then resuspended with 1 ml of the above-mentioned MES buffer solution, and then added 0.96 mg of EDC and 2.17 mg of NHS, Mix well, react at a reaction temperature of 37° C. for 0.5 hour, and then wash with a borax buffer solution with a concentration of 50 mmol and a pH of 8.5 to obtain 2.5 mg of activated magnetic particles.

[0065] Take 0.15 mg of anti-HBsAg antibody and the activated ma...

Embodiment 3

[0070] (1) Preparation of anti-HBsAg antibody labeled with superparamagnetic composite particles

[0071] Superparamagnetic Fe with a particle size of 300nm, a magnetic saturation intensity of 30emu / g, a corresponding external magnetic field response speed of 100 seconds, and a surface carboxyl content of 300μmol / g was used. 3 o 4 Nanoparticles.

[0072] The specific method is: take 2.5mg of the above-mentioned superparamagnetic Fe 3 o 4 Nanoparticles were washed with MES buffer solution with a concentration of 0.1 mol and a pH of 4.7, and separated and enriched with a 0.4T magnetic rack, then resuspended with 1 ml of the above-mentioned MES buffer solution, and then added 0.96 mg of EDC and 2.17 mg of NHS, Mix well, react at a reaction temperature of 36° C. for 20 minutes, and then wash with a borax buffer solution with a concentration of 50 mmol and a pH of 8.5 to obtain 2.5 mg of activated magnetic particles.

[0073] Take 0.15 mg of anti-HBsAg antibody and the activate...

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Abstract

The invention relates to an immunochromatographic test paper for detecting hepatitis B surface antigens and a preparation method thereof, belonging to the field of a hepatitis B surface antigen detection technique. The immunochromatographic test paper comprises a sample pad, a nitrocellulose membrane and a water absorption pad which are sequentially connected, wherein the sample pad contains a superparamagnetic composite particle labeled anti-HBsAg antibody; the nitrocellulose membrane is coated with a detection line and a quality control line which are mutually separate; the detection line contains an anti-HBsAg coating antibody; and the quality control line contains goat anti-mouse IgG which can be specifically combined with the anti-HBsAg coating antibody. The immunochromatographic test paper has the advantages of high sensitivity and high specificity, is quick and convenient, and can implement objective determination.

Description

technical field [0001] The invention belongs to the technical field of detection of hepatitis B surface antigen, and in particular relates to an immunochromatographic test paper for detecting hepatitis B surface antigen and a preparation method thereof. Background technique [0002] HBsAg, hepatitis B surface antigen, is the outer membrane protein of hepatitis B virus, and is the first HBV marker to appear in serum. In acute hepatitis, it disappears quickly. If it does not disappear after 6 months, it can become a carrier of chronic hepatitis (CH) or HBsAg, and it can last for several years or decades. The detection of HBsAg is of great significance for the diagnosis and differentiation of hepatitis B, epidemiological investigation, screening of blood donors, prognosis judgment, investigation of treatment effect and drug screening. [0003] Superparamagnetic composite particles have good magnetic properties and are especially suitable for the detection of biological samples...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/531
CPCG01N33/5764G01N33/558G01N2446/86
Inventor 马岚
Owner 昆明云大生物技术有限公司
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