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Human erythrocyte membrane antigen coated microsphere and application thereof

A technology of red blood cell membrane and cell membrane, which is applied in the application field of antibody detection, can solve the problems of short validity period, harsh storage conditions, unstable quality, etc., and achieve the effect of stable antigenicity, long validity period, and simple and easy method

Inactive Publication Date: 2011-01-26
INTEC PROD INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In order to solve this problem, people have adopted low-temperature freezing technology, but low-temperature freezing method is difficult to preserve a large number of red blood cells
Moreover, the antigenicity of red blood cells will decrease with the extension of frozen storage time.
In addition, the problem of difficult preservation of red blood cells also limits its application in other fields, such as quality control products for positive typing reagents, or anti-erythrocyte antibody titer determination and other fields
At present, fresh red blood cell reagents have problems such as difficulty in obtaining batches, short validity period, harsh storage conditions, and unstable quality.

Method used

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  • Human erythrocyte membrane antigen coated microsphere and application thereof
  • Human erythrocyte membrane antigen coated microsphere and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1: Preparation of red blood cell membrane antigen extracts for type A, type B and RhD positive blood groups

[0028] Take 1 ml of fresh A / B venous blood, centrifuge at 3000 rpm for 5 minutes to remove the supernatant, and collect the packed red blood cells for the next step. 10 ml of 5% sodium dodecyl sulfonate (SDS) was added to the packed red blood cells, and after standing at room temperature for 3 minutes, the supernatant was removed by centrifugation at 3000 rpm. 10 ml of pure water was added to the precipitate, mixed by shaking, and centrifuged again at 3000 rpm to remove the supernatant. This is repeated 5 times, and the obtained precipitate is the A / B type erythrocyte membrane antigen extract.

[0029] Take 1 ml of fresh RhD-positive venous blood and centrifuge at 3000 rpm for 5 minutes to collect cell pellets. Add 10 ml of sodium dodecyl sulfonate (SDS) to the packed red blood cells. After standing at room temperature for 3 minutes, the supernatant w...

Embodiment 2

[0030] Example 2: Preparation of red-dyed polystyrene-acrylic microspheres

[0031] Take 70ml of absolute ethanol and add 30ml of pure water, mix well and set aside. To the obtained aqueous ethanol were sequentially added 1.0 g of ammonium persulfate, 10 ml of styrene and 5 ml of acrylic acid. After mixing, high-purity nitrogen gas was introduced (10 minutes). Then, it was placed in a 60° C. water bath and stirred, and the reaction was terminated after continuous heating for 48 hours to obtain polystyrene-acrylic acid microspheres.

[0032]Take the polystyrene-acrylic microspheres prepared above, and centrifuge to remove the supernatant. 100 ml of pure water was added to the precipitate, and 1 g of sodium dodecyl sulfonate (SDS) was added. Take another vial and add 10 ml of absolute ethanol, 0.01 g of methyl red, 1 ml of styrene, and 0.03 g of azoisobutyronitrile. Dissolve and shake well and add to the liquid mixture prepared above. The reaction was completed after heatin...

Embodiment 3

[0033] Example 3: Preparation of RhD-positive erythrocyte membrane antigen-coated silica microspheres

[0034] 100 ml of absolute ethanol was added to 10 ml of ethyl orthosilicate and 10 ml of ammonia water, and the reaction was completed after stirring at room temperature for 24 hours to obtain silica microspheres. Take 20 ml of the silica microspheres thus prepared, centrifuge to remove the supernatant, and leave the precipitate.

[0035] The RhD-positive type O erythrocyte membrane antigen prepared in Example 1 was added to the silica microsphere precipitate, the pH value was adjusted to 10, the mixture was mixed and shaken moderately. The supernatant was removed by centrifugation to obtain RhD-positive erythrocyte membrane antigen-coated silica microsphere precipitate.

[0036] 10 ml of 10 mg / ml Rhodamine B solution was added to the above RhD positive erythrocyte membrane antigen-coated silica microsphere pellet. After staining at 37°C for 2 hours, bright red RhD-positiv...

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Abstract

The invention provides a method for preserving the activity of a human cell membrane blood group antigen, which comprises the steps of: preparing an erythrocyte membrane blood group antigen extract, and then coating the prepared erythrocyte membrane blood group antigen on a solid microsphere so as to replace a fresh erythrocyte to be used for detecting a blood group antibody in a sample.

Description

Field of Invention [0001] The present invention relates to a blood group detection material and a method for long-term maintenance of erythrocyte antigen activity, in particular to human erythrocyte membrane antigen-coated microspheres and their application in the detection of human blood group antibodies. Background of the Invention [0002] The research and detection of human cell membrane antigens and their specific antibodies is one of the most basic contents of biological science, immunological experimental research and clinical detection. Human ABO and Rh blood group antigen / antibody detection are routine clinical detection items. Blood typing is based on cell agglutination by antigen-antibody reactions. The use of red blood cell surface antigens to detect blood group antibodies in serum is called reverse typing. In clinical blood transfusion, it is necessary to ensure that the identification results of positive and negative stereotyping methods are consistent to ens...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N33/551G01N33/80G01N33/531G01N33/546
Inventor 江应玲曹大与汪大明钟乾兴肖江群
Owner INTEC PROD INC
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