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Kit for detecting coupling efficiency of target antibody and microspheres as well as method and application thereof

A technology for coupling efficiency and target detection, applied in biological testing, measuring devices, material inspection products, etc., can solve the problems of long time consumption, low detection sensitivity of protein detection, cumbersome operation, etc.

Pending Publication Date: 2022-01-14
上海艾瑞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

ELISA method is highly sensitive and specific for the detection of residual antibodies in the supernatant, but it is time-consuming, cumbersome and has many influencing factors
However, the Bradford method has a large deviation in the determination of different proteins; the BCA method is not sensitive to the detection of proteins and takes a long time

Method used

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  • Kit for detecting coupling efficiency of target antibody and microspheres as well as method and application thereof
  • Kit for detecting coupling efficiency of target antibody and microspheres as well as method and application thereof
  • Kit for detecting coupling efficiency of target antibody and microspheres as well as method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] A method for detecting the coupling efficiency of a target antibody and microspheres, comprising the following steps.

[0041] (1) Preparation of Immunolateral Flow Chromatography Reagent Tablets

[0042] Preparation of nitrocellulose membrane:

[0043] Preparation of T-line (detection line 31) drawing solution: dilute a goat anti-mouse IgG (detection line antibody) to 0.5mg / mL with pH 7.3, 0.01M PBS diluent, and set aside at 2-8°C;

[0044] Preparation of line C (quality control line 32) drawing solution: dilute any mouse monoclonal antibody (quality control line antibody) to 2 mg / mL with pH 7.3, 0.01M PBS diluent, and store at 2-8°C for later use.

[0045] Scribe the T-line and C-line scribing fluids on the nitrocellulose membrane (NC membrane) with a scribing and gold-spraying instrument, and then dry the NC membrane for 2 hours in a vacuum to obtain a test line 31 and a quality control line. 32 nitrocellulose membrane.

[0046] (2) Detection of antibody-coupled m...

Embodiment 2

[0053] A method for detecting the coupling efficiency of antibodies and microspheres, roughly the same as in Example 1, the difference mainly lies in the difference between the detection line antibody, the quality control line antibody and the target antibody, the differences are as follows:

[0054] The T line is coated with an antigen (D-Dimer, in other embodiments, it can also be antigens such as SAA and CRP), and the C line is coated with any mouse monoclonal antibody to detect a mouse monoclonal antibody (D-Dimer). Dimer mouse monoclonal antibody, in other embodiments, can be SAA, CRP and other mouse monoclonal antibodies) labeling efficiency on fluorescent microspheres.

[0055] Preparation of T-line drawing solution: Dilute an antigen (such as D-Dimer, SAA, CRP and other antigens) to 0.5 mg / mL with pH 7.3, 0.01M PBS diluent, and store it at 2-8°C for later use.

Embodiment 3

[0057] A method for detecting the coupling efficiency of antibodies and microspheres is roughly the same as in Example 1, with the differences as follows:

[0058] Preparation of T-line drawing solution: dilute a goat anti-mouse IgG (detection line antibody) to 1, 0.75 or 0.5mg / mL with pH 7.3, 0.01M PBS diluent, and set aside at 2-8°C;

[0059] Preparation of C-line marking solution: dilute any mouse monoclonal antibody (quality control line antibody) to 4, 3 or 2 mg / mL with pH 7.3, 0.01M PBS diluent, and store at 2-8°C for later use.

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Abstract

The invention discloses a kit for detecting the coupling efficiency of a target antibody and microspheres as well as a method and application thereof, and relates to the technical field of immunochromatography detection. The method comprises the following steps: adding a conjugate of a detection antibody and a microsphere into a supernatant to form a to-be-detected substance, capturing a residual target antibody after coupling of a target antibody and the microsphere, then detecting the to-be-detected substance by using immunochromatography test paper, and capturing a compound formed by the microsphere, the detection antibody and the target antibody by using a detection line; enabling a quality control line to capture a conjugate formed after the microsphere and the detection antibody are coupled, and acquiring the amount of the residual target antibody in the supernate by detecting the detection line and the quality control line to obtain the coupling efficiency of the target antibody and the microsphere. The method for detecting the coupling efficiency of the target antibody and the microspheres has the advantages of simplicity, convenience and rapidness in operation, low cost and good accuracy, and a novel way is provided for detecting the coupling efficiency of the antibody and the microspheres.

Description

technical field [0001] The invention relates to the technical field of immunochromatography detection, in particular to a kit for detecting the coupling efficiency of a target antibody and microspheres, a method and an application thereof. Background technique [0002] The immunological lateral flow chromatography technology is designed based on the principle of immunology, and uses the antigen-antibody immunological reaction to realize the qualitative or quantitative detection of the substance to be tested through the specific combination between the antigen and the antibody. The main principle is: the antigen or antibody labeled with microspheres reacts with the object to be detected, and then binds to the specific ligand coated on a solid-phase carrier such as nitrocellulose membrane to form lines or spots. The instrument can achieve the purpose of qualitative or quantitative analysis of the markers to be detected by reading or measuring these lines or spots. [0003] Am...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/558G01N33/68G01N33/543
CPCG01N33/558G01N33/6854G01N33/54313
Inventor 成晶陈超马亚飞
Owner 上海艾瑞德生物科技有限公司
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