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Coupling method of immune magnetic particles, diluent and application

A technology of immunomagnetic particles and diluents, which is applied in the biological field, can solve the problems of poor dispersion, poor chemiluminescence test performance, and low coupling efficiency of immunomagnetic particles, and achieve reduced interference, good dispersion and stability of magnetic beads, The effect of improving coupling efficiency

Active Publication Date: 2021-08-27
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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  • Coupling method of immune magnetic particles, diluent and application
  • Coupling method of immune magnetic particles, diluent and application
  • Coupling method of immune magnetic particles, diluent and application

Examples

Experimental program
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Effect test

Embodiment 1

[0039] Example 1: Preparation of immunomagnetic particles

[0040] Step 1: Activation of carboxyl magnetic beads

[0041] (1) Take 10mg of magnetic beads, wash and incubate with 1ml 100mM MES (pH5.0) for 2min, absorb on a 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 take an appropriate amount of diluted EDC and add it to the magnetic beads in step (3). The mass ratio of EDC:magnetic beads is 1:1, and vortex to mix Evenly, incubate at room temperature for 30 min;

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

[0045] (5) Wash once with 100mM MES (pH5.0), absorb on a magnetic plate, and discard the supernatant;

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

[0047] Step 2: Antibody-magnetic bead coupling

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

Embodiment 2

[0054] Embodiment 2: Preparation of immunomagnetic particle dilution

[0055] The components of the immunomagnetic particle 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 analyzer, and the RLU values ​​were recorded.

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Abstract

The invention provides a coupling method of immune magnetic particles, a diluent and application. According to an antibody-magnetic bead coupling process, carboxyl magnetic beads are activated under an acidic condition, then the activated carboxyl magnetic beads are coupled to an antibody under a nearly neutral condition, and under the mild reaction condition, the biological activity of the antibody is kept, aggregation of the activated carboxyl magnetic beads is reduced, and the efficiency of coupling the carboxyl magnetic beads to the antibody is improved. According to the confining liquid provided by the invention, carboxyl sites, which are not coupled to an antibody, on the magnetic beads are combined by BSA, amino acid or / and small molecule peptides and PEG through ionic bonds or covalent bonds, and meanwhile, the antibody coupled to the carboxyl magnetic beads are kept in a correct and stable form, so that the detection performance is improved, and the sensitivity is enhanced. According to the immune magnetic particle diluent provided by the invention, the prepared immune magnetic particles, such as CA125 antibody-carboxyl magnetic beads, can still keep good dispersity and stability after being diluted by the diluent and placed at 4 DEG C for 12 months or at 37 DEG C for 7 days.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a coupling method, dispersion liquid and application of immunomagnetic particles. Background technique [0002] Magnetic particles are widely used in chemiluminescent immunoassays. The common surface active groups include carboxyl and amino groups. Under certain conditions, they can be activated by intermediates and can be coupled with amino or carboxyl groups on antibodies or antigens for use in antigens. or antibody detection. There are one-step method and two-step method for common carboxyl magnetic beads coupling antibody: the one-step method is to mix the magnetic beads and antibody, and then add a cross-linking agent to react. 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 will often be affected, so this cross-linking method is...

Claims

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Application Information

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