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High gradient magnetic separation of biological material

A biological material, magnetic separation technology, applied in high gradient magnetic separation, high gradient magnetic separator, magnetic separation and other directions, can solve problems such as non-specific binding, achieve simple cost and improve the effect of purification results

Active Publication Date: 2011-02-16
X ZELL INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The solution of the present invention is based on the principle that the separation of non-target particles from the HGMS separation column is solved by a buffer solution for equilibrating the separation column and for suspending the biological material to be separated, having the properties stated in the claims. The problem of non-specific binding of the matrix

Method used

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  • High gradient magnetic separation of biological material
  • High gradient magnetic separation of biological material
  • High gradient magnetic separation of biological material

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0072] Using isotonic phosphate-buffered sucrose solution containing gelatin against Plasmodium falciparum (P.falciparum) cultures of red blood cells infected with the malaria pathogen (Plasmodium) purification.

[0073] Material:

[0074] buffer solution A 1 : Isotonic phosphate-buffered sucrose solution containing 0.75% gelatin

[0075] Stainless steel wool 1g

[0076] single tap

[0077] three way tap

[0078] 20G injection needle

[0079] 3ml disposable syringe

[0080] 10ml disposable syringe

[0081] 50ml disposable syringe

[0082] 1 neodymium horseshoe magnet

[0083] a) Preparation of purification kit

[0084] Manufacture of Separation Columns

[0085] A 3 ml disposable syringe used as separation column 1 was filled with 1 gram of stainless steel wool as matrix 2 to two-thirds of the total volume of the 3 ml disposable syringe. Here, it should be noted that a large number of stainless steel wool fibers are located in the longitudinal direction of the ...

example 2

[0098] Use phosphate-buffered saline solution (PBS) containing bovine serum albumin (BSA) Purification of malaria pathogens (Plasmodium) from cultures of P. falciparum infected red blood cells

[0099] Material:

[0100] As listed in Example 1, but with Buffer A 1 buffered solution A 2 (PBS with 5% BSA) instead.

[0101] a) Preparation of purification kit

[0102] Follow the description in Example 1.

[0103] b) Preparation and execution of the separation process

[0104] Preparation of P. falciparum cultures for purification of red blood cells infected with the malarial pathogen

[0105] As in Example 1. The parasitemia of the P. falciparum culture in this experiment was 14.47%. buffer solution A 1 with buffer solution A 2 replace.

[0106] Execution of the separation process

[0107] As in Example 1, with the following differences:

[0108] buffer solution A 1 with buffer solution A 2 replace. Centrifugation of the eluate was then performed at 800 g for ...

example 3

[0112] Purified from a suspension of leukocytes (peripheral blood mononuclear cells (PBMCs)) Original CD8 leukocytes (CD8 positive cells).

[0113] Material:

[0114] as listed in Example 1.

[0115] a) Preparation of purification kit

[0116] Follow the description in Example 1.

[0117] b) Preparation and execution of the separation process

[0118] Labeling of CD8-positive cells with antibody-conjugated synthetic paramagnetic particles (microbeads)

[0119] will be 1.5x10 7 Individual human peripheral mononuclear cells (PBMCs) were incubated with monoclonal rat anti-human CD8 IgG antibody in PBS / BSA 1% for 30 minutes on ice. The cells were washed twice with the same buffer solution, and then incubated with anti-rat IgG-coupled microbeads (MiltenyiBiotech GmbH, loc. cit.) for an additional 10 minutes on ice. The cells were washed twice in the same buffer solution, and then with fluorescein-labeled antibodies (PE-anti-CD8 antibody and FITC-anti-CD3 antibody, Simulte...

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Abstract

The invention relates to a high gradient magnet separation device for the separation or purification of magnetic or magnetically labeled biological material, comprising a magnet (7), a separating column (1), and a ferromagnetic matrix (2) that can be arranged in an interior chamber of the separating column, and a storage container (11) containing a buffer solution for the equilibration of the separating column (1) and / or for the suspension of the biological material, wherein during operation a magnetic field created by the magnet (7) creates a high gradient magnetic field in the matrix (2) and buffer solution can flow from the storage container (11) through the separating column (1). For this purpose, the buffer solution comprises a base solution and macromolecules that can saturate nonspecific binding sites of the matrix (2).

Description

technical field [0001] The present invention relates to the application of high gradient magnetic separation (HGMS) technology for the separation and purification of biological material. Background technique [0002] Especially in the field of biomedical research, the separation and purification of defined particles from heterogeneous particle suspensions is extremely important for various analytical methods. In general, the particles to be purified, hereinafter referred to as "target particles", are usually only minimally different from the rest of the particles contained in the suspension, hereinafter referred to as "non-target particles" . Target particles and non-target particles are typically cells or cell fragments, but can also be any other biological material. [0003] Some existing separation methods exploit the magnetic properties of target particles, where the target particles have naturally occurring "intrinsic" magnetic properties, or where the target particle...

Claims

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Application Information

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IPC IPC(8): B03C1/28B03C1/01
CPCB03C1/01B03C2201/18B03C1/288B03C1/032B03C1/0335B03C1/034B03C2201/26B03C1/002C12N13/00
Inventor 塞巴斯蒂安·恰克里特·巴克蒂普里达·玛拉西特
Owner X ZELL INC
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