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Kit and use of SPA (staphylococcal protein A)-antibody trimer

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

Inactive Publication Date: 2013-10-23
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

[0044] Example 1: Cell Treatment Kit

[0045] 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.

[0046] 2. Human lymphocyte separation medium (commercially available) 100ml, centrifugal density 1.077g / ml.

[0047] 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

[0051] Example 2: Cell Treatment Kit

[0052] 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.

[0053] 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.

[0054] 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

[0058] Example 3: Spa-antibody

[0059] 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, and has also 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 technique that can be easily applied at the grassroots level.

[0060] 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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Abstract

The invention relates to a kit and use of SPA (staphylococcal protein A)-antibody trimer. The antibody trimer is formed by an anti erythrocyte antibody, SPA and antibodies of some anti-leukocyte antigen or other antigens. According to the antibody trimer, the self anti erythrocyte antibody is combined with erythrocyte, the other antibody is combined with corresponding leukocyte antigen (or other antigen), then leukocyte (or other cell, factor) and erythrocyte are deposited by a blood sedimentation process or methods, such as density gradient centrifugation to achieve the purpose of eliminating components of corresponding cell and the like, and the erythrocyte can be also connected with corresponding antigen by virtue of the SPA to be used as means for detecting certain antigens. The kit is convenient and simple and can be directly used for clinical separation and extraction of stem cells / progenitor cells, and the collected and extracted stem cells / progenitor cells do not have any foreign markers. The kit based on the trimer can be industrially produced to realize popularization of human hematopoietic stem cell separation and purification technology.

Description

[0001] The present invention is a divisional application for an invention patent titled "SPA-antibody trimer, a cell treatment kit containing the trimer, its preparation method and use" (application number 201010022753.4, application date January 13, 2010) patent. technical field [0002] 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 cell treatment kit and its application. technical background [0003] 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%. [0004] SPA and IgG with co-precipitation reaction must have...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0789
Inventor 张厚亮
Owner 王信
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