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Immunomagnetic bead for separating red blood cells, and preparation method and application of immunomagnetic bead

An immunomagnetic bead, red blood cell technology, applied in cell dissociation methods, biochemical equipment and methods, animal cells, etc., can solve problems such as unfavorable rapid detection, rupture of blood cells, and long time-consuming static separation, so as to reduce hemolysis, Avoid hemolysis, easy to achieve effect

Inactive Publication Date: 2020-09-15
北京康思润业生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The main purpose of the present invention is to provide a kind of immunomagnetic beads for separating red blood cells and its preparation method and application. The technical problem to be solved is that the existing separation method takes too long to stand and separate, and improper centrifugation will cause blood cells The rupture is not conducive to rapid detection

Method used

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  • Immunomagnetic bead for separating red blood cells, and preparation method and application of immunomagnetic bead
  • Immunomagnetic bead for separating red blood cells, and preparation method and application of immunomagnetic bead

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preparation example Construction

[0031] One embodiment of the present invention proposes a method for preparing immunomagnetic beads for separating red blood cells, which includes:

[0032] Step S1, wash the magnetic beads, remove impurities, add phosphate buffer solution to the washed magnetic bead precipitation, make the pH value in the range of 5.0-6.5, preferably pH 6.0, acidic conditions are more conducive to the activation of groups, ultrasonic mixing Homogenize the bead solution, the purpose of ultrasonic is to mix evenly, the preferential ultrasonic condition is 40kHz, 6min, add activator to the magnetic beads, and ultrasonically mix, the preferential ultrasonic condition is 40kHz, 6min, the combination of the magnetic beads and activator The weight ratio is 1:0.1-5, and then the activated solution is placed on a shaker, and shaken and reacted at 37° C. at 100 rpm for 24 minutes.

[0033] Step S2, adding the anti-erythrocyte antibody to the activated product obtained in step S1, and performing ultraso...

Embodiment 1

[0065] A preparation method for immunomagnetic beads for separating red blood cells, specifically comprising the following steps:

[0066] 1.1.1 Washing of magnetic beads:

[0067] The original concentration of the magnetic beads was 2.0wt%, adding 10mM phosphate buffer (pH 6.0), centrifuging with a strong magnetic field, discarding the supernatant, and repeating twice to obtain the magnetic beads precipitate.

[0068] 1.1.2 Activation:

[0069] 1) Prepare the activator solution:

[0070] The activator is N-hydroxysuccinimide and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide with a weight ratio of 1:1, respectively using 10mM (pH 6.0) phosphate buffer solution to prepare a 500mg / mL solution for later use;

[0071] 2) Activation of magnetic beads:

[0072] Add 10mM phosphate buffer solution (pH 6.0) to the washed magnetic bead pellet, and ultrasonically mix (40kHz, 6min) to obtain a 100mg / mL magnetic bead solution. Mixed solution of succinimide and 1-ethyl-3-(3-dimethylam...

Embodiment 2

[0084] The immunomagnetic beads obtained in Example 1 are used to separate red blood cells in plasma. When in use, take 0.5 mL of whole blood and add it to the vial in Example 4, shake well, and then place it on a magnetic stand or a magnet. After 2 minutes, you can obtain upper plasma.

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Abstract

The invention relates to an immunomagnetic bead for separating red blood cells, and a preparation method and application of the immunomagnetic bead, and relates to the technical field of preparation of immunomagnetic beads. The preparation method of the immunomagnetic bead comprises the following steps: S1, activating a magnetic bead with an activating agent, wherein a weight ratio of the magneticbead to the activating agent is 1:(0.1-5), and an activation time is 5-30 min; S2, adding an anti-red blood cell antibody into the activated product obtained in the step S1, and carrying out ultrasonic treatment; and S3, adding a sealing agent into the product obtained in the step S2 for sealing, and performing strong magnetic separation to obtain the immunomagnetic bead for separating red bloodcells. The immunomagnetic bead obtained by the method disclosed by the invention is used for separating red blood cells in blood plasma, the separation of the blood cells can be completed within 1-2 min, the time is saved, a basis is provided for the rapid detection, and meanwhile, the hemolysis phenomenon caused by factors such as centrifugation is avoided, so that the accuracy of a detection result is improved.

Description

technical field [0001] The invention belongs to the technical field of preparation of immunomagnetic beads, and in particular relates to an immunomagnetic bead for separating red blood cells, a preparation method and application thereof. Background technique [0002] Immunomagnetic separation (IMS) is a method for cell sorting that has emerged in recent years. It coats the surface of magnetic beads with immunoreactive antibodies for antigen-antibody reaction. Form an "antigen-antibody-magnetic bead" immune complex. Once these bead-bound cells are placed under a powerful magnetic field, they move in a directed manner, sorting the immune complexes from other unbound cells. When the superparamagnetic magnetic beads leave the magnetic field, the magnetism disappears immediately, so as to achieve the purpose of positive or negative selection of specific cells. [0003] The examination of blood components in clinical examination requires the separation of serum or plasma in whol...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/078G01N33/543
CPCC12N5/0641G01N33/54326C12N2509/10
Inventor 张军李海燕徐学晗宁妍郭海鹏
Owner 北京康思润业生物技术有限公司
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