A composition for lysing red blood cells, red blood cell lysing reagent and application
A red blood cell lysing and reagent technology, which is applied in the field of lysing red blood cell compositions, can solve the problems of poor compatibility of lysing reagents, lack of formula, poor performance, etc., and achieve low red blood cell lysing efficiency and sample compatibility, increase permeability, improve The effect of controllability
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Embodiment 1
[0048] Embodiment 1 is used for the preparation general method of the composition of erythrocyte lysis and liquid reagent
[0049] The present embodiment provides a method for preparing a composition for erythrocyte lysis and a liquid reagent, comprising the following steps:
[0050] Under the condition that the temperature is 20-25℃ and the humidity is less than 60%, use a precision balance of 1 / 10,000 or 1 / 100,000 to accurately weigh the solid or liquid material of the corresponding mass, and place it in a 50mL sterile centrifuge. In a tube, the composition for erythrocyte lysis is obtained;
[0051] The above composition is fully dissolved in water for injection of corresponding quality to form a colorless and transparent solution;
[0052] In a sterile environment with local 100-level laminar flow purification conditions, the obtained solution was filtered through a 0.22 μm diameter filter membrane into another 50 mL sterile centrifuge tube to obtain a liquid reagent fo...
Embodiment 2
[0053] Embodiment 2 is used for the composition of the liquid reagent of erythrocyte lysis
[0054] This example provides a set of liquid reagents for erythrocyte lysis, the composition of which is shown in Table 1. Among them, the liquid medium numbered LA01, LA02, LA03, LA04, LA05, LA06, LA07, LA08, LA09, LA10, LA11, LA12, LA13, LA14, LA15 is the liquid medium provided by the present invention for processing biological samples- - Red blood cell lysis reagent, liquid reagents numbered LA16, LA17, LA18 are comparative examples, where LA18 means water for injection without any solute.
[0055] Table 1 Composition of liquid reagents (by weight percentage, the balance is water for injection)
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Embodiment 3
[0058] Example 3 Using the control group and the liquid reagent of the present invention to lyse red blood cells in human peripheral blood and evaluate the lysing effect
[0059] Human peripheral blood samples were taken from healthy adult volunteers, volunteers were between 20-40 years old, regardless of gender;
[0060] Use commercially available vacuum blood collection tubes to collect peripheral blood, the vacuum blood collection tubes are pre-filled with anticoagulant, and the anticoagulant is selected from EDTAK2, sodium citrate or heparin;
[0061] The commercially available lysis reagent product was used as the experimental control group to illustrate the use effect of the lysis reagent of the present invention, including control group A (RBCLysisBuffer, Biolegend, USA), control group B (ACKlysingbuffer, Lonza, Switzerland), control group C (hemolyzing agent for flow cytometry, Kuangbo Biological Co., Ltd.); density gradient medium (Ficoll-Paque TM PLUS, GE Healthca...
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Abstract
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