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Cell sorting method by using avidin/streptavidin magnetic composite particles

A technology of composite particles and avidin, applied in blood/immune system cells, bone/connective tissue cells, animal cells, etc., can solve the problems of low antibody binding, non-specific protein adsorption, low cell purity and efficiency, etc. To achieve the effect of low cytotoxicity, reduction of non-specific adsorption problems, and simple and time-saving operation

Active Publication Date: 2010-09-29
XIAN GOLDMAG NANOBIOTECH
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Problems solved by technology

However, there are still some problems in the current magnetic composite particles used in cell sorting. For example, the magnetic composite particles have the disadvantages of non-specific adsorption of proteins other than the target antibody, or the low binding amount of antibodies, resulting in the purity of the target cells sorted in cell sorting. and inefficient

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  • Cell sorting method by using avidin/streptavidin magnetic composite particles
  • Cell sorting method by using avidin/streptavidin magnetic composite particles

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Embodiment

[0030] Example: Positive sorting of peripheral blood mononuclear cells (PBMC) using avidin / streptavidin magnetic composite particles (the target cell is CD3).

[0031] The sample cells to be sorted are: use lymphocyte separation medium to extract peripheral blood mononuclear cells (PBMC) from anticoagulated blood, or prepare single cell suspension from lymphoid tissue. Add a certain amount of PBS (5% heat-inactivated fetal bovine serum, pH = 7.4, 0.09% NaN 3 ) Shake well, turn at 5000 for 5min, repeat the above operation and wash 3 times, filter through 50um nylon mesh to remove cell clumps and debris, adjust the number of cells to 1×10 7 .

[0032] (1) Pretreatment of avidin / streptavidin magnetic composite particles

[0033] Disperse the avidin / streptavidin magnetic composite particles in a balanced salt solution, the balanced salt solution is: pH=7.0, 1M tris-hydrochloric acid solution (Tris-HCl), magnetically separated, discarded clear;

[0034] (2) closed

[0035] Add...

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Abstract

The invention relates to a method for sorting target cells by using avidin / streptavidin magnetic composite particles. In the method, a biotin-labeled monoclonal antibody is added into an experimental system, magnetic particles are bonded with the biotin-labeled monoclonal antibody through avidin / streptavidin, the monoclonal antibody is specifically bonded with corresponding antigen on the surfaceof the cells to make the cells captured by the magnetic particles so as to purify and separate the cells. Because the biotin-avidin and biotin-streptavidin have the characteristics of high system specificity, high stability, wide application range and low experiment cost, the obtained avidin / streptavidin magnetic composite particles have high biotin bonding property; and the labeled amount is improved in the measurement technique, and biomolecules are difficult to denature or inactivate, so that the cell sorting specificity is further improved. The high-purity target cells can be sorted by specifically bonding the biotin with the avidin or streptavidin; and the method has the advantages of no need of expensive instrument, simple operation and time saving.

Description

technical field [0001] The invention relates to the field of biological detection, in particular to a method for sorting target cells by using avidin / streptavidin magnetic composite particles. Background technique [0002] With the continuous development of life sciences, the basic research of cell biology and immunology and the diagnosis of clinical diseases all need to separate purified cells from mixed samples containing different cells. The existing cell separation technologies mainly include: Ficoll density gradient centrifugation, Centrifugal washing, two-phase separation, these methods are based on the characteristics of physical properties such as cell size, density, surface charge and so on. High purity and recovery require expensive instruments, and the operation process is complicated and time-consuming. The use of magnetic composite particles for cell sorting is a cell separation technology that has developed rapidly in recent years. It uses magnetic composite p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/078C12N5/077C12N5/0786
Inventor 崔亚丽高启祥石峰丁海琴陈超杨柳
Owner XIAN GOLDMAG NANOBIOTECH
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