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A coupling method, diluent and application of immunomagnetic particles

An immunomagnetic particle and coupling technology, which is applied in the biological field, can solve the problems of poor chemiluminescence test performance, low coupling efficiency of immunomagnetic particles, and poor dispersion, and achieve good dispersion and stability of magnetic beads and improve coupling Efficiency, the effect of reducing interference

Active Publication Date: 2022-05-27
NINGBO RUI BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The technical problem to be solved by the present invention is to provide a coupling method, dispersion liquid and application of immunomagnetic particles to solve the problem of low coupling efficiency and poor dispersion of immunomagnetic particles in the prior art and the use of immunomagnetic particles to prepare chemiluminescence assays. poor performance

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  • A coupling method, diluent and application of immunomagnetic particles
  • A coupling method, diluent and application of immunomagnetic particles
  • A coupling method, diluent and application of immunomagnetic particles

Examples

Experimental program
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Effect test

Embodiment 1

[0039] Example 1: Preparation of Immunomagnetic Microparticles

[0040] Step 1: Activation of carboxyl magnetic beads

[0041] (1) Take 10 mg of magnetic beads, wash with 1 ml of 100 mM MES (pH 5.0) and incubate for 2 min, adsorb on the magnetic plate, and discard the supernatant;

[0042] (2) Add 100mM MES (pH5.0) to resuspend;

[0043] (3) Take 100mM MES (pH5.0) to dissolve EDC to 10mg / ml, and add an appropriate amount of diluted EDC to the magnetic beads in step (3). The mass ratio of EDC:magnetic beads is 1:1, and vortex mixing Homogenize and incubate at room temperature for 30 min;

[0044] (4) After the incubation, the magnetic plate is adsorbed, and the supernatant is discarded;

[0045] (5) Wash once with 100 mM MES (pH 5.0), adsorb on the magnetic plate, discard the supernatant;

[0046] (6) Add 100mM Good's buffer (pH6.8) to prepare activated carboxyl magnetic beads;

[0047] Step 2: Antibody-Magnetic Bead Conjugation

[0048] The CA125 antibody was diluted wit...

Embodiment 2

[0054] Example 2: Preparation of Immunomagnetic Microparticle Diluent

[0055] The components of the immunomagnetic microparticle dilution include: 0.1M Tris (pH7.5), 200mM NaCl, 1% BSA (w / v), 5% PVP (w / v), 2% trehalose (w / v), 0.05% Tween 20 (v / v), 0.05% Sodium Azide, 0.05% ProClin300.

[0056] The CA125 antibody-magnetic beads were diluted 1:10 (v / v) with the above diluent and used as a solid-phase reagent. The background values ​​were detected on an automatic chemiluminescence immunoassay, and the RLU values ​​were recorded.

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Abstract

The invention provides a coupling method, diluent and application of immunomagnetic particles. The antibody-magnetic bead coupling process of the present invention first activates the carboxyl magnetic beads under acidic pH conditions, and then couples the activated carboxyl magnetic beads with the antibody under near-neutral conditions. The mild reaction conditions maintain The biological activity of the antibody also reduces the aggregation of the carboxyl magnetic beads after activation, and improves the efficiency of the carboxyl magnetic bead-coupled antibody. The blocking solution provided by the present invention enables the carboxyl sites of the uncoupled antibody on the magnetic beads to be ionically or covalently bound by BSA, amino acids or / and small molecule peptides and PEG, and at the same time makes the carboxyl sites coupled to the carboxyl magnetic beads Antibodies maintain their correct and stable form, improving assay performance and enhancing sensitivity. The immunomagnetic particle diluent provided by the present invention can make the prepared immunomagnetic particles, such as CA125 antibody-carboxyl magnetic beads, after being diluted with the diluent, still keep good after being placed at 4°C for 12 months or at 37°C for 7 days dispersion and stability.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a coupling method, dispersion and application of immunomagnetic particles. Background technique [0002] Magnetic particles are widely used in chemiluminescence immunoassays. Common surface active groups include carboxyl and amino groups. Under certain conditions, through intermediate activation, they can be coupled with amino or carboxyl groups on antibodies or antigens for antigens. or antibody detection. Common carboxyl magnetic beads conjugated antibody methods include one-step method and two-step method: one-step method is to mix magnetic beads and antibody, and then add cross-linking agent for reaction. The one-step method has higher requirements for antibodies and needs to tolerate higher concentrations of cross-linking agents. However, if the cross-linking dose is too low, the coupling efficiency is often affected, so this cross-linking method is rarely used. [0003] The t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/543G01N33/574
CPCG01N33/54326G01N33/57484
Inventor 陈沈雪陈媛张闻
Owner NINGBO RUI BIO TECH