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Method for rapidly separating supernatant from human whole blood or suspended red blood cells by utilizing lyophilized preparation

A technology of freeze-dried preparations and red blood cells, which is applied in the field of clinical testing and medical scientific research, can solve problems such as time-consuming, meet different applications and needs, accelerate the speed of magnetic separation, and facilitate popularization and application.

Inactive Publication Date: 2009-06-03
NANJING GENERAL HOSPITAL NANJING MILLITARY COMMAND P L A +2
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  • Summary
  • Abstract
  • Description
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  • Application Information

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Problems solved by technology

[0002] Separating the supernatant of whole blood or suspended red blood cells is one of the most commonly used techniques in clinical and scientific research. The commonly used separation methods include natural sedimentation and centrifugation. The former takes a long time, while the latter requires the guarantee of a centrifuge and electricity

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  • Method for rapidly separating supernatant from human whole blood or suspended red blood cells by utilizing lyophilized preparation

Examples

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Effect test

Embodiment 1

[0029] Example 1: Method for Separating Whole Blood or Suspended Red Blood Cell Supernatant Using Phytohemagglutinin Pre-coated Magnetic Particles

[0030] Take 100g of Chixiaodou, wash it with pure water, soak it in 200ml of pure water for 24 hours, use a juicer to crush the Chixiaodou into soybean milk, add 50ml of pure water during the crushing process to improve the grinding efficiency, and centrifuge the milk at 6000g for 30 Minutes, the supernatant was retained and further purified by conventional saturated ammonium sulfate method to obtain pure plant lectin for future use.

[0031] According to the ratio of pre-coating 20 μg of phytohemagglutinin per milliliter of 10% concentration of magnetic polymer microspheres (average diameter 100nm) solution, the magnetic polystyrene microspheres were dissolved in PBS (50mM, pH7.4) buffer in advance, The concentration of the microspheres in the solution is 10% (m / v), the lectin is added under the condition of rapid stirring, and t...

Embodiment 2

[0035] Example 2: Method for Separating Suspended Red Blood Cell Supernatant Using Anti-erythrocyte Antibody Pre-coated Magnetic Particles

[0036] Take 1 mg of anti-erythrocyte antibody purchased from Beijing Xinchuang Bioengineering Co., Ltd. According to the magnetic polymer microspheres of 10% concentration per milliliter, the average diameter of the microspheres is 100nm, and the solution is pre-coated with 20 μg of anti-erythrocyte antibody ratio, the magnetic polymer microspheres are dissolved in PBS buffer (50mM pH7.4) in advance, The concentration of the microspheres in the buffer was 10% (m / v), and the anti-erythrocyte antibody was added under the condition of rapid stirring, and the reaction was stirred at a speed of 100 rpm at room temperature for 30 minutes, and BSA was added to the final concentration of BSA in the solution. The concentration is 0.2%, and the reaction is stirred at 100 rpm at room temperature for 15 minutes. Finally, the supernatant is discarded ...

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Abstract

The invention relates to a method for rapidly separating supernatant from human whole blood or suspended red blood cells by utilizing lyophilized preparation. The method comprises the following steps: the phytoagglutinin or the anti erythrocyte antibody is pre-coated on a magnetic polymer microsphere to be prepared into isolating reagent for the liquid whole blood or suspended red blood cells; the isolating reagent for the liquid whole blood or suspended red blood cells is absorbed by insoluble polymer, and after lyophylization, the isolating reagent is cut into thin slices to be prepared into isolating reagent for lyophilized whole blood or suspended red blood cells; and the isolating reagent for the lyophilized whole blood or suspended red blood cells is added to the human whole blood or suspended red blood cells to be uniformly mixed and then placed above a magnetic body, and the supernatant is rapidly separated. The lyophilized preparation preserved for long term at the normal temperature can be prepared by the method, and the supernatant can be rapidly separated form the human whole blood or suspended red blood cells under the conditions without ensuing the electric power and the centrifugal machine.

Description

technical field [0001] The present invention relates to a method for rapidly separating human whole blood or suspended erythrocyte supernatant using a freeze-dried preparation, in particular to a method for separating human by using freeze-dried polymer microspheres coated with phytohemagglutinin or anti-erythrocyte antibody and containing magnetic materials. Methods for supernatants from whole blood or suspended erythrocytes. The invention can be widely used in the fields of clinical examination, medical scientific research and the like. Background technique [0002] Separating the supernatant of whole blood or suspended red blood cells is one of the most commonly used techniques in clinical and scientific research. The commonly used separation methods include natural sedimentation and centrifugation. The former takes a long time, while the latter requires the guarantee of centrifuges and electricity. Contents of the invention [0003] The purpose of the present inventio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/08C12N5/078
Inventor 栾建凤王言帅丁万和朱培元
Owner NANJING GENERAL HOSPITAL NANJING MILLITARY COMMAND P L A
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