Immunomagnetic bead for peripheral blood lymphocyte separation and preparation and application thereof

An immunomagnetic bead and lymphocyte technology, applied in the field of nanomaterials, can solve the problems of easy agglomeration, low adsorption capacity, increased sorting cost, etc., and achieve the effects of excellent biocompatibility, controllable synthesis process, and simple preparation.

Inactive Publication Date: 2017-02-01
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

First of all, the stability of immunomagnetic nanoparticles is generally insufficient, and they are pr...

Method used

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  • Immunomagnetic bead for peripheral blood lymphocyte separation and preparation and application thereof
  • Immunomagnetic bead for peripheral blood lymphocyte separation and preparation and application thereof
  • Immunomagnetic bead for peripheral blood lymphocyte separation and preparation and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] A preparation method for immunomagnetic beads for peripheral blood lymphocyte separation, comprising the following steps:

[0043] (1) Preparation of one-step carboxylated magnetic nanoparticles:

[0044]Mix ferric citrate, ferrous sulfate solution and polyacrylic acid additive in a molar ratio of 0.5:0.5:0.01, stir evenly with magnetic force at room temperature, then slowly add sodium hydroxide solution to adjust the pH value to 8, hydrothermal reaction for 8 hours, The temperature is 180°C. After the reaction is completed, dialyze with a dialysis bag with a molecular weight cut-off of 12K for 1 day to obtain a dispersed and purified MNPs-COOH liquid;

[0045] (2) Preparation of specific immunomagnetic beads:

[0046] ①Activation of carboxyl groups by magnetic nanoparticles:

[0047] Take 500 microliters of MNPs-COOH purified solution, centrifuge with 500 microliters of ultrafiltration tubes with a molecular weight cut-off of 10K, and remove the supernatant. The cond...

Embodiment 2

[0056] A preparation method for immunomagnetic beads for peripheral blood lymphocyte separation, comprising the following steps:

[0057] (1) Preparation of one-step carboxylated magnetic nanoparticles:

[0058] Mix ferric citrate, ferrous chloride solution and carboxymethyl cellulose additive in a molar ratio of 0.8:1:0.01, stir evenly with magnetic force at room temperature, then slowly add sodium hydroxide solution dropwise to adjust the pH value to 9, water Heat the reaction for 10 hours at 200°C. After the reaction is completed, dialyze with a dialysis bag with a molecular weight cut-off of 12K for 1 day to obtain the MNPs-COOH dispersion and purification solution;

[0059] (2) Preparation of specific immunomagnetic beads:

[0060] ①Activation of carboxyl groups by magnetic nanoparticles:

[0061] Take 500 microliters of MNPs-COOH purified solution, centrifuge with 500 microliters of ultrafiltration tubes with a molecular weight cut-off of 10K, remove the supernatant, a...

Embodiment 3

[0070] A preparation method for immunomagnetic beads for peripheral blood lymphocyte separation, comprising the following steps:

[0071] (1) Preparation of one-step carboxylated magnetic nanoparticles:

[0072] Mix iron dextran, ferrous sulfate solution and alginic acid additive in a molar ratio of 1.5:0.5:0.03, stir evenly with magnetic force at room temperature, then slowly add sodium hydroxide solution to adjust the pH value to 12, hydrothermal reaction for 12 hours, temperature After the reaction is completed, use a dialysis bag with a molecular weight cut-off of 14K to dialyze for 3 days to obtain MNPs-COOH dispersion and purification liquid;

[0073] (2) Preparation of specific immunomagnetic beads:

[0074] ①Activation of carboxyl groups by magnetic nanoparticles:

[0075] Take 500 microliters of MNPs-COOH purified solution, centrifuge with 500 microliters of ultrafiltration tubes with a molecular weight cut-off of 10K, remove the supernatant, and set the conditions ...

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PUM

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Abstract

The invention relates to an immunomagnetic bead for peripheral blood lymphocyte separation. The immunomagnetic bead comprises carboxylic ferroferric oxide magnetic nanoparticles and a specific antibody, wherein the mass ratio of the carboxylic ferroferric oxide magnetic nanoparticles to the specific antibody is (2-5) to (0.1-0.5), a specific antibody is selected from one or more of CD8, CD3E, IL2RA and CD4. The immunomagnetic bead is obtained by preparing the carboxylic ferroferric oxide magnetic nanoparticles uniform in particle size distribution, good in water solubility and excellent in magnetic property through a hydrothermal method and then coupling the nanoparticles with the specific antibody. Compared with the prior art, the immunomagnetic bead has the advantages that the bead is simple in preparation process and good in stability, a matched separation column is large in sample amount, high in positive selection rate, low in cost and the like, sufficient high-activity lymphocytes can be obtained, and the lymphocytes can be preserved for revival, culture and re-delivery at a proper time.

Description

technical field [0001] The invention relates to the field of nanomaterials, in particular to an immunomagnetic bead used for separating lymphocytes in peripheral blood and its preparation and application. Background technique [0002] Immunological micro-nanosphere technology is a technology based on immunology, using various micro-nanospheres covalently coupled with immunologically active substances for immunological or other biological analysis and separation. Since 1956, the technology has emerged in various fields such as immunology, microbiology, biochemistry, molecular genetics and molecular biology because of its simplicity, efficiency and practicality. [0003] Immunomagnetic micronanospheres or immunomagnetic micronanobeads for short, as a new type of functional material, has been widely studied in recent years, and its application in the fields of biomedicine and clinical diagnosis has attracted great attention from researchers from all over the world. Immunomagne...

Claims

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Application Information

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IPC IPC(8): C07K17/14C12N5/078B82Y5/00B82Y25/00B82Y30/00B82Y40/00
CPCC07K17/14B82Y5/00B82Y25/00B82Y30/00B82Y40/00C12N5/0634C12N2509/00
Inventor 高国尹婷张倩黄鹏崔大祥
Owner SHANGHAI JIAO TONG UNIV
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