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SPA-antibody tripolymer, cell treating kit containing tripolymer, preparation method and application thereof

A trimer and antibody technology, applied in the field of biomedicine, can solve the problems of high cost, high technical requirements, and difficulty in clinical promotion and application, and achieve the effect of easy promotion, simple method, and low cost

Inactive Publication Date: 2014-07-16
王信
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The common problems of these methods are: firstly, all types of stem cells cannot be effectively preserved; secondly, the cost is expensive and the technical requirements are relatively high
There are certain difficulties in clinical application

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1: Cell Treatment Kit

[0043] 1. Trimer of SPA (staphylococcal protein A), red blood cell antibody and specific lymphocyte antibody. The preparation process of the trimer: the reaction concentrations of SPA and anti-erythrocyte monoclonal antibody and anti-cd3 antibody, cd14 antibody, cd19 antibody and cd56 antibody are all 0.1 mg / ml. SPA is 1.5ml, anti-erythrocyte monoclonal antibody is 1ml, anti-cd3 antibody, cd14 antibody, cd19 antibody, and cd56 antibody are each 0.25ml. Incubate in a 37°C water bath for 30 minutes. Add PBS solution, adjust the concentration of Spa-antibody trimer to 1 μg / ml, volume to 100ml, pH 7.4. Store at 4°C.

[0044] 2. Human lymphocyte separation medium (commercially available) 100ml, centrifugal density 1.077g / ml.

[0045] 3. Cell chromatography plate: polystyrene chromatography plate, after disinfection and sterilization, add anti-cd3 antibody, cd14 antibody, cd19 antibody, and cd56 antibody at a concentration of 0.1mg / ml, just ...

Embodiment 2

[0049] Example 2: Cell Treatment Kit

[0050] 1. Trimer of SPA (staphylococcal protein A), red blood cell antibody and specific lymphocyte antibody. The preparation process of the trimer: the reaction concentrations of SPA and anti-erythrocyte monoclonal antibody and anti-cd3 antibody, cd14 antibody, cd19 antibody and cd56 antibody are all 0.1 mg / ml. SPA is 1.5ml, anti-erythrocyte monoclonal antibody is 1ml, anti-CD3 antibody, CD14 antibody, CD19 antibody, CD56 antibody is 0.25ml. Incubate in a 37°C water bath for 30 minutes. Add PBS to make the Spa-antibody trimer concentration 1 μg / ml, volume 100ml, pH 7.4. Store at 4°C.

[0051] 2. Lymphocyte separation solution 100ml: an aqueous solution composed of 0.3% sodium diatrizoate and 8% polysucrose 400#, with a density of 1.077g / ml.

[0052] 3. Antibody conjugate: mix anti-CD3 antibody, CD14 antibody, CD19 antibody, CD56 antibody and nanoparticles of polystyrene material. The antibody concentration is 0.1 mg / ml, the volume is...

Embodiment 3

[0056] Example 3: Spa-antibody

[0057] Chicken Marek's disease (MD) is a highly contagious lymphoproliferative disease caused by Marek's virus. Its main features are: lymphocyte infiltration and swelling of peripheral nerves, various organs, iris and skin. Marek's disease mainly occurs in chickens, but also has been reported in turkeys, pheasants, ducks, quails, swans and other poultry. The disease is highly contagious, spreads quickly and has a long incubation period. Acute onset chickens are eliminated and the mortality rate reaches 10% to 80%. So far, in the diagnosis of Marek's disease, except for AGPT, there is no practical technology that can be easily applied at the grassroots level.

[0058] Trimer of SPA (staphylococcal protein A) and erythrocyte antibody, anti-Marek's virus antibody. The preparation process of the trimer: the reaction concentrations of SPA, anti-erythrocyte monoclonal antibody and anti-Marek's virus antibody are all 0.1 mg / ml. SPA is 1.5ml, ant...

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PUM

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Abstract

The invention relates to an SPA-antibody tripolymer, a cell treating kit containing the tripolymer, a preparation method and application thereof. The tripolymer comprises an anti erythrocyte antibody, an SPA and an antibody of a certain anti leukocyte antigen or other antigens. The tripolymer is combined with an erythrocyte through the anti erythrocyte antibody per se, the other antibody is combined with a corresponding leukocyte antigen (or other antigens), and then leukocyte (other cells, factors) and the erythrocyte are deposited through an erythrocyte sedimentation process or other methods of density gradient centrifugation and the like, thereby achieving the purpose of eliminating ingredients of corresponding cells and the like, connecting the erythrocyte with a corresponding antigen by the SPA, and being used as a mean for detecting a certain antigen. The invention has convenience and simpleness, and can be directly used for clinically separating and extracting stem cells / progenitor cells; and the collected and extracted stem cells / progenitor cells have no external markers. Meanwhile, the kit based on the tripolymer can be industrially produced so as to realize the popularization of the separation and purification technology of hematopoietic stem cells.

Description

technical field [0001] The invention relates to the field of biomedical technology, in particular to a staphylococcal protein A (staphylococal protein A, hereinafter referred to as SPA)-antibody trimer, a cell treatment kit based on the polymer, and a preparation method and use thereof. technical background [0002] SPA can bind to Fc fragments in human and various mammalian serum IgG molecules. The order of binding affinity is pig, dog, rabbit, human, monkey, rat, mouse and cow. In addition to binding to IgG, SPA can also bind to a small amount of IgM and IgA in serum. The adsorption rate of SPA bacteria to various immunoglobulins: 90% to 98% for IgG, 2% to 30% for IgM, and 1.5% for IgA %~20%. [0003] SPA and IgG with co-precipitation reaction must have two binding sites, which are respectively on the Fc and Fab segments. Although the lack of one can bind to SPA, co-precipitation reaction does not occur. The participation of this Fab is not SPA-antibody reaction. Studie...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N33/577G01N33/544C12N5/0789
Inventor 张厚亮
Owner 王信
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