Immunomagnetic composite, preparation method thereof, antigen capture reagent, kit and system comprising same, and applications

A complex and magnetic technology, applied in the field of biomedicine, can solve the problems of reducing the activity of lectins, and achieve the effect of increasing the coupling capacity, ensuring biological activity, and reducing steric hindrance

Inactive Publication Date: 2018-07-27
SHENZHEN NEW INDS BIOMEDICAL ENG
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AI Technical Summary

Problems solved by technology

[0010] The present invention aims to provide an immunomagnetic complex, its preparation method, its antigen capture agent, kit, system and application to solve the problem of agglutination caused by direct coupling of lectins on the surface of magnetic microspheres in the prior art. The direct contact between the lectin and the rigid interface of the magnetic ball reduces the technical problem of lectin activity

Method used

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  • Immunomagnetic composite, preparation method thereof, antigen capture reagent, kit and system comprising same, and applications

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preparation example Construction

[0042] According to a typical embodiment of the present invention, a method for preparing any one of the above immunomagnetic complexes is provided. The preparation method comprises the following steps: 1) using amino polymer to modify magnetic microspheres to obtain amino functionalized magnetic microspheres; 2) linking lectin to amino polymers to form immunomagnetic complexes.

[0043] Amino polymer coated magnetic microspheres make the surface of the magnetic microspheres modified with a large number of amino groups. The outer polymer has a dendritic network structure, which can fully increase the coupling capacity, reduce steric hindrance, and make the target coupling The lectin is far away from the rigid surface of the magnetic microspheres, which ensures the biological activity of the target conjugate to the greatest extent. When the immunomagnetic complex is used to capture and separate AFP-L3, its capture and separation efficiency will be greatly improved.

[0044] Pr...

Embodiment approach

[0060] According to a typical embodiment of the present invention, a kit for capturing, separating and quantitatively detecting AFP-L3 is provided. The kit includes any one of the above-mentioned antigen capture agents.

[0061] Preferably, the kit also includes: a low concentration calibrator, a high calibrator, an anti-AFP antibody (preferably an anti-AFP monoclonal antibody) labeled with a luminescent marker, and a buffer; preferably, the concentration of the low concentration calibrator is 10.936- 20.313IU / Ml, the concentration of the height calibrator is 350-650IU / Ml; preferably, the working concentration of the luminescent marker is 5ng / mL-500ng / mL, and the working concentration of the anti-AFP antibody is 50ng / mL-5000ng / mL; preferably Yes, the buffer is carbonate buffer, phosphate buffer, borate buffer or Tris-HCl buffer; preferably, the luminescent marker is luminol, isoluminol and derivatives thereof; alkaline Phosphatase, horseradish peroxidase, fluorescent substanc...

Embodiment 1

[0096] A method for enhancing the capture and separation efficiency of AFP-L3 and its detection kit

[0097] Preparation 1: Preparation of lentil lectin oxidized modification

[0098] Dissolve 2 mg of lentil lectin with freshly prepared 0.2 mol / L sodium bicarbonate aqueous solution, add freshly prepared 0.5 mL 8 mmol / L sodium periodate aqueous solution dropwise to the above solution, keep away from light, and slowly The reaction was stirred for 2 hours to obtain the oxidized product of lentil lectin (LECTIN-CHO).

[0099] Preparation 2: Preparation of polyethyleneimine-mediated magnetic microspheres

[0100] Take a certain amount of magnetic balls, add 0.5mg of DCC per milligram of magnetic balls, then add ethanol solution to make the concentration of magnetic balls 20mg / mL, place in a water bath at 38°C and shake or mechanically stir for 2 hours; the activation is good Remove the supernatant from the nano-magnetic sphere solution, and then add carbonate buffer solution of p...

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Abstract

The invention discloses an immunomagnetic composite, a preparation method thereof, an antigen capture reagent, a kit and a system comprising same, and applications. The immunomagnetic composite includes: amino-functionalized magnetic microspheres formed by modifying magnetic microspheres with an amino polymer, and agglutinin jointed with the amino polymer. By means of the technical scheme, the agglutinin is actually connected to the magnetic microspheres through induction by the amino polymer coating the surfaces of the magnetic microspheres, wherein the surfaces of the magnetic microspheres are modified by a large quantity of amino groups by coating the magnetic microspheres with the amino polymer. The outer polymer has a dendritic network structure, so that conjugation load is fully increased, and steric hindrance is reduced. In addition, the target conjugate, agglutinin, is far away from the rigid surface of the magnetic microspheres, so that bioactivity of the target conjugate is guaranteed as most as possible.

Description

technical field [0001] The present invention relates to the technical field of biomedicine, in particular, to an immunomagnetic complex, a preparation method thereof, an antigen capture agent comprising the same, a kit, a system and an application thereof. Background technique [0002] Primary liver cancer (primary hepatic carcinoma, PHC) is a common malignant tumor, and its mortality rate ranks third in all tumors, second only to lung cancer and gastric cancer. The number of people who die of liver cancer in my country accounts for 45% of the world's deaths every year . In recent years, it is generally believed that the occurrence of PHC is the result of long-term effects of multiple factors, multiple channels, and multiple steps, including external environmental carcinogenic factors (parasites, bacteria, virus infection, aflatoxin intake, water pollution, smoking, drinking, etc.) ) and hereditary factors. Early diagnosis of PHC is particularly important because of its ins...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/531
CPCG01N33/531
Inventor 饶微刘坤袁锦云刘来壮李婷华陈曙光
Owner SHENZHEN NEW INDS BIOMEDICAL ENG
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