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Two-well immunoassay lateral flow test card

A technology of immunological lateral chromatography and test card, which is applied in the field of new double-hole immunological lateral chromatography test card, which can solve problems such as reagent failure, tailing, and instability of gold particles, and achieve the effect of improving accuracy

Active Publication Date: 2019-01-11
天津东亚生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Gold particles prepared by gold standard technology are often not uniform in diameter, and gold particles with non-uniform diameters move at different speeds during chromatography, which leads to tailing
In addition, the gold particles are unstable and easy to coagulate or precipitate, leading to reagent failure
Third, the method of labeling proteins is cumbersome

Method used

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  • Two-well immunoassay lateral flow test card

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1: Labeling HLA-G monoclonal antibody A1 with red polystyrene microspheres

[0037] (1) Prepare colored polystyrene microspheres: adopt commercially available red polystyrene microspheres with a diameter of 10-150 nanometers, with active carboxyl or amino groups on them;

[0038] (2) HLA-G monoclonal antibody A1 was dialyzed overnight with pH8.0-9.0, 0.01mol / L tris-HCL buffer, and then centrifuged to remove the modified protein precipitate for later use;

[0039] (3) Suspend polystyrene colored microspheres in pH8.0-9.0, 0.01mol / L tris-HCL buffer solution, in protein cross-linking agent carbodiimide (EDC), N-hydroxysulfosuccinyl In the presence of imine, adipic dihydrazide, etc., add HLA-G monoclonal antibody A1 dropwise under constant stirring; continue stirring for 10 minutes after adding HLA-G monoclonal antibody A1, then add PEG20000 to make PEG20000 in the buffer The final concentration in the solution reaches 0.1% to stabilize the colored immune microsphe...

Embodiment 2

[0042] Embodiment 2: Preparation of novel dual-hole immunological lateral flow test card (referred to as test card)

[0043] (1) Prepare labeled antibodies A1, A2, and A3 of HLA-G, CEA, and HBsAg. Set the labeled antibodies A1 and A2 on the glass fiber membrane near well A, and set the labeled antibody A3 on the glass fiber membrane near well B. Please refer to the attached figure 1 .

[0044] (2) Prepare the capture antibodies B1, B2, and B3 for HLA-G, CEA, and HBsAg. Fix the capture antibodies B1 and B2 on the NC membrane at the T1 and T2 marks of the A hole window. Immobilize the capture antibody B3 on the NC membrane at the T3 mark in the window of well B. Goat anti-rabbit IgG is immobilized at the mark C of the window of well B.

[0045] (3) Holes A and B of the test card are the holes where the whole blood from the tip is dripped.

Embodiment 3

[0046] Embodiment 3: The principle of using a double-hole test card to detect HLA-G:

[0047] Assuming that a blood sample only contains HLA-G, HBsAg, and does not contain CEA, the chromatographic process is demonstrated as follows:

[0048] When the blood sample to be tested is dropped into hole A and hole B of the test card at the same time, in hole A, the red-labeled antibody A1 of HLA-G, which is set on the glass fiber membrane near hole A, combines with the HLA-G antigen in the sample to form HLA - G-A1 red immune complex. This red immune complex moves forward along the NC membrane. When reaching the T1 mark, the HLA-G capture antibody B1 immobilized at the T1 mark binds to HLA-G in the HLA-G-A1 red immune complex to form a red line. It indicates the presence of HLA-G in the blood sample. The HLA-G-A1 red immune complex continues to move forward. When passing the T2 mark, the HLA-G-A1 red immune complex does not react with the CEA capture antibody immobilized on the T2...

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Abstract

The invention discloses a novel double-hole immunity lateral chromatography test card. An antibody of a broad-spectrum tumor marker HLA-G (Human Leukocyte Antigen-G) is used as a main substance, an antibody of a carcino-embryonic antigen (CEA) and an antibody of a hepatitis B surface antigen (HBsAg) are taken as reference substances, the HLA-G is jointly detected by three antibodies to form the novel double-hole immunity lateral chromatography test card for screening cancer. By using the novel double-hole immunity lateral chromatography test card to detect HLA-G, false positive and false negative results can be eliminated, and the accuracy of a detection result is improved. According to the novel double-hole immunity lateral chromatography test card disclosed by the invention, colored polymer microspheres are utilized to replace gold particles, so that particles of a colloidal solution are more uniform and stable.

Description

technical field [0001] The invention relates to the technical field of biomedical clinical immune analysis, in particular to a novel double-hole immunological lateral flow chromatography test card. Background technique [0002] Tumor is a common disease and frequently-occurring disease, and its incidence rate ranks second. According to the report of the World Health Organization in 1997, the annual death toll of cancer patients in the world is as high as 6.6 million, and by 2020, the number of cancer patients in the world will reach 15 million. Various malignant tumors seriously threaten human health. [0003] At present, researchers have discovered a variety of markers related to malignant tumors, such as AFP (alpha-fetoprotein), CEA (carcinoembryonic antigen), CA15-3, CA19-9, CA50, CA125, PSA and so on. However, these tumor-associated markers are monofunctional markers. The use of monofunctional tumor markers can only detect the corresponding type of tumor, but it can't...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/574G01N33/558
CPCG01N33/558G01N33/57484
Inventor 苏殿杰
Owner 天津东亚生物技术有限公司
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