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Gelatinized immunomagnetic bead as well as preparation method and application thereof

A technology of immunomagnetic beads and gelation, which is applied in the direction of chemiluminescence/bioluminescence, analysis and measurement devices through chemical reactions of materials, and can solve the problems of self-crosslinking of antibodies, serious aggregation of immunomagnetic beads, and antibody grafting. It can improve the hydrophilicity, reduce the self-crosslinking of antibodies, and achieve good suspension.

Active Publication Date: 2020-12-04
WUHAN LIFE ORIGIN BIOTECH LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Comparative Document 1: Publication No. "CN108508195A", titled "An Immunomagnetic Bead and Its Preparation Method and Application" discloses an organogel-type immunomagnetic bead synthesized by the glutaraldehyde method. The preparation method utilizes metal -The nature of organogel-immobilized proteins. Immobilizing antibodies and ferric oxide in a metal-organic gel solid system at the same time is simple and convenient, but the prepared immunomagnetic beads are likely to cause self-crosslinking of antibodies and grafting of antibodies Low rate, severe aggregation of immunomagnetic beads

Method used

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  • Gelatinized immunomagnetic bead as well as preparation method and application thereof
  • Gelatinized immunomagnetic bead as well as preparation method and application thereof
  • Gelatinized immunomagnetic bead as well as preparation method and application thereof

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preparation example Construction

[0038] According to a typical embodiment of the present invention, a preparation method of gelled immunomagnetic beads is provided, such as figure 2 shown, including:

[0039] S1, reacting amino magnetic beads and gelatin under the action of a dianhydride organic acid anhydride initiator to obtain magnetic gelatin microspheres;

[0040] S2, using NHS solution, EDC solution and AEM to activate the magnetic gelatin microspheres to obtain a maleamidated magnetic gelatin microsphere solution;

[0041] S3, obtaining an activated antibody solution;

[0042] S4. Perform a coupling reaction between the maleamidated magnetic gelatin microsphere solution and the activated antibody solution to obtain a coupling reaction solution;

[0043] S5. Block the coupling reaction solution, then perform magnetic separation to obtain a solid, and resuspend the solid with a storage buffer to obtain gelled immunomagnetic beads.

[0044] A kind of gelling immune magnetic bead provided by the presen...

Embodiment 1

[0083] The chemiluminescence immunoassay kit in this embodiment includes R reagent and M reagent.

[0084] R reagent is the working solution prepared in advance, and the main component is alkaline phosphatase-labeled CTNI 1.

[0085] M reagent preparation process: Dilute the gelled immunomagnetic beads obtained in the embodiment of the present invention with M buffer solution into a working solution for later use, specifically:

[0086] (1) Solution preparation

[0087] Cross-linking buffer A: 100 mM pH 8.5 boric acid buffer.

[0088] Cross-linking buffer B: 100 mM pH 7.0 HEPES buffer.

[0089] Activator: use DMF (N,N dimethylformamide) to configure 20mg / ml initiator butane carboxylic acid dianhydride; use crosslinking buffer B to prepare 50mg / mL Sulfo-NHS and EDC·HCl; Configure 40mg / mL AEM in linking buffer B; configure 10mg / mL Traut's reagent in cross-linking buffer B. All activators need to be prepared and used immediately.

[0090] Blocking solution: 10% BSA.

[0091...

Embodiment 2

[0101] In Example 1, the initiator butane carboxylic dianhydride was replaced with cyclopentane tetracarboxylic dianhydride; the antibody was replaced with NT-proBNP2, and other components and processes were the same as in Example 1.

[0102] Control group 1

[0103] According to the synthesis method of immunomagnetic beads in CN108508195A, the antibody is replaced by CTNI2 antibody, and other processes and components remain unchanged.

[0104] Control group 2

[0105] According to the synthesis method of immunomagnetic beads in CN108508195A, the antibody is replaced by NT-proBNP2 antibody, and other processes and components remain unchanged.

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Abstract

The invention discloses a gelatinized immunomagnetic bead as well as a preparation method and application thereof. The preparation method comprises the following steps that: amino magnetic beads and gelatin react under the action of a dianhydride organic anhydride initiator to obtain magnetic gelatin microspheres; the magnetic gelatin microspheres are activated by using an NHS solution, an EDC solution and an AEM to obtain a maleamidated magnetic gelatin microsphere solution; an activated antibody solution is obtained; coupling reaction is performed on the maleamidated magnetic gelatin microsphere solution and the activated antibody solution to obtain a coupling reaction solution; and the coupling reaction solution is sealed, then magnetic separation is performed to obtain a solid, and thesolid is resuspended by using a preservation buffer solution to obtain gelatinized immunomagnetic beads. The gelatinized immunomagnetic beads provided by the invention can improve the grafting rate of an antibody and retain the activity of the antibody; and the immunomagnetic beads are applied to a chemiluminescence immunoassay project, so that the sensitivity and stability of the project are improved.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a gelled immunomagnetic bead and its preparation method and application. Background technique [0002] Immunomagnetic beads are a functional complex composed of magnetic microspheres and immune proteins. After immunomagnetic beads are specifically immunized with the target, the target can be separated from the mixed solution under the action of a magnetic field. Based on the above characteristics, immunomagnetic beads are widely used as an important component in chemiluminescence immunoassay. The hydrophilic property of the magnetic microspheres used as the carrier of the immunomagnetic beads will affect the anti-non-specific adsorption performance of the immunomagnetic beads, and at the same time, its suspension performance will affect the stability of the immunomagnetic beads. [0003] Gelatin is a chain with a variety of active functional groups (-NH 2 , -COOH, -OH, etc.) water...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/531G01N33/543G01N21/76
CPCG01N33/531G01N33/54333G01N33/5434G01N21/76G01N2446/84G01N2446/86Y02A50/30
Inventor 王铎来祥兵李重阳舒芹张雪娇
Owner WUHAN LIFE ORIGIN BIOTECH LTD
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