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A kind of erythrocyte membrane and its preparation method and application
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A technology of erythrocyte membrane and erythrocytes, applied in the fields of medicine and biology, can solve the problem of not having rare blood type antigens, and achieve the effect of ensuring the safety of blood transfusion
Active Publication Date: 2020-03-13
GENERAL HOSPITAL OF PLA
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[0005] One object of the present invention is to provide a red blood cell membrane with rare blood group antigenicity on the surface of the red blood cell membrane, which solves the problem that the existing red blood cell membrane usually only has ABO blood group antigenicity but not rare blood group antigenicity
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Embodiment 1
[0030] Embodiment 1 prepares erythrocyte membrane
[0031] (1) Collect packed red blood cells, add an equal volume of normal saline, quickly freeze in liquid nitrogen at -196°C, and then thaw in a water bath at 37°C;
[0032] (2) Repeat the operation of "freezing-thawing" 8 times;
[0033] (3) Centrifuge at 15,000 g for 20 minutes, remove the supernatant, and obtain a white precipitate that is the red blood cell membrane.
Embodiment 2
[0034] Embodiment 2 prepares erythrocyte membrane
[0035] (1) Collect packed red blood cells, add an equal volume of normal saline, freeze at -40°C, and then melt in a 37°C water bath;
[0036] (2) Repeat the operation of "freezing-thawing" 3 times;
[0037] (3) Centrifuge at 15000g for 20min, remove the supernatant;
[0038](4) Washing with pure water for 5 times, the obtained white precipitate is red blood cell membrane;
[0039] (5) The homogenate was crushed for 20 minutes;
[0040] (6) Observe the morphology and particle size under a microscope;
[0041] (7) Filter with a filter membrane with a pore size of 5 μm;
[0042] (8) Determination of the red blood cell membrane concentration after filtration in step (7) with a BCA kit.
Embodiment 3
[0043] Embodiment 3 prepares erythrocyte membrane
[0044] (1) Take packed red blood cells, add half volume ratio of normal saline, freeze at -80°C, and then thaw at room temperature of 20°C;
[0045] (2) Repeat the "freezing-thawing" operation 5 times.
[0046] (3) centrifuge at 5000g for 30min, remove the supernatant;
[0047] (4) Washing with pure water for 10 times, the obtained white precipitate is red blood cell membrane;
[0048] (5) The homogenate was crushed for 40 minutes;
[0049] (6) Use a filter membrane to filter after observing the morphology and particle size under a microscope;
[0050] (7) Determination of concentration with BCA kit.
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Abstract
The invention provides an erythrocyte membrane, and a preparation method and application thereof. A repeated freeze-thaw process is further combined with a homogenization process and filter membrane filtration to prepare the erythrocyte membrane which has the advantages of uniform particle size distribution and favorable antigenicity (particularly favorable antigenicity for the rare blood group systems. The erythrocyte membrane solves the problem that the erythrocyte membrane in the prior art usually only has the ABO blood group antigens and has no rare blood group antigens, and also solves the problem of nonuniform particle size distribution in the existing erythrocyte membrane. The invention provides an erythrocyte membrane which has uniform particle size distribution and has whole bloodgroup system antigens, and can be used for screening and identification of unexpected antibodies, thereby ensuring the blood transfusion safety.
Description
technical field [0001] The invention belongs to the technical field of medicine and biology, and relates to the preparation of red blood cell membranes, in particular to a red blood cell membrane and its preparation method and application. Background technique [0002] The main components of the cell membrane are proteins (about 49% by weight), lipids (about 44% by weight, mainly including phospholipids, cholesterol, glycolipids, etc.) and sugars, which have important biological functions, such as material metabolism and energy conversion. , cell recognition, cellular immunity, and tumorigenesis are all highly correlated, and have extensive research and application value in the fields of molecular biology, cellbiology, immunology, and clinical medical testing. For the human body, the number of red blood cells is large, and the material is easy to obtain, and the mature red blood cells of the human body have no nucleus and organelles. Due to the imbalance of osmotic pressure...
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