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Immune capture composition, preparation method, kit and application

An immunocapture and composition technology, applied in the field of immunodetection, can solve the problems of high background signal, affecting the sensitivity and specificity of detection results, and poor blocking effect.

Active Publication Date: 2020-07-07
SHENZHEN DYMIND BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The main technical problem to be solved in this application is to provide an immunoassay composition, preparation method, kit and application, which can solve the problem of protein as a single blocking agent due to blocking The effect is not good, resulting in high background signal, which affects the sensitivity and specificity of the detection results

Method used

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  • Immune capture composition, preparation method, kit and application
  • Immune capture composition, preparation method, kit and application
  • Immune capture composition, preparation method, kit and application

Examples

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preparation example Construction

[0054] refer to figure 1 , the application provides a preparation method of an immunodetection composition, the preparation method comprising the following steps:

[0055] S101: Provide a solid phase carrier.

[0056] Specifically, the solid phase carrier is a microwell plate, a glass sheet, a microfluidic chip, latex, polymer microspheres, metal nanoparticles, metal alloy nanoparticles, inorganic microspheres, inorganic / organic hybrid microspheres, fluorescent nanoparticles At least one of magnetic beads, multifunctional fluorescent magnetic beads; metal nanoparticles include but not limited to gold nanoparticles or silver nanoparticles. Preferably, the solid phase carrier is polymer microspheres.

[0057] Commercial carboxyl-modified superparamagnetic microspheres can be used as the magnetic bead carrier. After washing with buffer, 1-ethyl-3-(3-dimethylaminopropyl)- The carbodiimide solution is shaken and reacted at room temperature for 30-60 minutes in the dark, and then...

Embodiment 1

[0103] Immunological detection composition 1 comprises the following components:

[0104] (1) Solid phase carrier: surface carboxylated magnetic beads with a diameter of 1-10 μm;

[0105] (2) Ligand: N-terminal BNP capture antibody;

[0106] (3) The first blocking compound: trehalose;

[0107] (4) The second blocking compound: bovine serum albumin.

[0108] The preparation of immunodetection composition 1 comprises the following steps:

[0109] Step (1): Wash 1mg of surface carboxylated magnetic beads (hereinafter referred to as magnetic beads) twice with MES buffer (50mM, pH6). 100 μg each of imide (NHS) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC), wherein 50 mM MES buffer at pH 6 was used for NHS and EDC Dissolve and activate at room temperature for 30 minutes. Wash twice with PBS buffer (10 mM, 0.1% Tween20) for magnetic separation. Add 40 μg of N-terminal BNP capture antibody (dissolved in PBS buffer) and react at room temperature for 2 hour...

Embodiment 2

[0115] Immunological detection composition 2 comprises the following components:

[0116] (1) Solid phase carrier: surface carboxylated magnetic beads with a diameter of 6 μm;

[0117] (2) Ligand: N-terminal BNP capture antibody;

[0118] (3) The first blocking compound: bovine serum albumin;

[0119] (4) The second blocking compound: trehalose.

[0120] The preparation of immunodetection composition 2 comprises the following steps:

[0121] Step (1): Wash 1mg of surface carboxylated magnetic beads (hereinafter referred to as magnetic beads) twice with MES buffer (50mM, pH6). 100 μg each of imide (NHS) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC), wherein 50 mM MES buffer at pH 6 was used for NHS and EDC Dissolve and activate at room temperature for 30 minutes. Wash twice with PBS buffer (10 mM, 0.1% Tween20) for magnetic separation. Add 40 μg of N-terminal BNP capture antibody (dissolved in PBS buffer) and react at room temperature for 2 hours. ...

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Abstract

The invention discloses an immunodetection composition, a preparation method, a kit and application. The immune capture composition comprises: a solid-phase carrier; the at least one first ligand thatis fixed on the surface of the solid-phase carrier; the first blocking compound and the second blocking compound that are modified on the surface of the solid-phase carrier and are used for blockingactive sites on the surface of the solid-phase carrier, wherein the first blocking compound is a polyhydroxy saccharide compound or a protein compound, the second blocking compound is a polyhydroxy saccharide compound or a protein compound, and the first blocking compound is different from the second blocking compound. By means of the mode, the sealing effect can be effectively improved, and the accuracy of the detection result is improved.

Description

technical field [0001] This application relates to the technical field of immune detection, in particular to immune capture composition, preparation method, kit and application. Background technique [0002] In the classical immunoassay method, high affinity and specific recognition between antigens and antibodies forms the basis of the analysis. Affinity is a general term for a series of non-covalent forces (hydrophobic, electrostatic, hydrogen bonds, etc.) between ligands and receptors under a specific biomolecular structure. The combined effect of multiple weak forces ensures the stable combination of both sides . However, non-target substances existing in complex systems may adsorb affinity ligands and receptors to varying degrees, which is called "non-specific adsorption". In actual experimental operation and related device design, it is usually necessary to add a blocking agent to bind to the active site of the sensing interface to reduce non-specific adsorption with...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N33/533G01N33/532
CPCG01N33/54393G01N33/533G01N33/532Y02A50/30
Inventor 马志亚钱婷汪业红李会平刘治志
Owner SHENZHEN DYMIND BIOTECH
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