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Method for screening respiratory syncytial virus memory B cells by nano magnetic bead process

A technology of nano-magnetic beads and syncytial virus, applied in animal cells, vertebrate cells, blood/immune system cells, etc., to achieve high sensitivity, simple operation, and rapid separation

Inactive Publication Date: 2015-04-29
NANCHANG UNIV
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Problems solved by technology

[0004] The purpose of the present invention is to provide a method for preparing nano-magnetic beads to screen respiratory syncytial virus memory B cells for the existing monitoring of RSV infection history and the shortcomings of RSV antibody preparation methods

Method used

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Embodiment Construction

[0011] The preparation method of the present invention includes the following steps:

[0012] 1. Connect the nanomagnetic beads and the GD and GL antigen proteins: add 20ul each of the respiratory syncytial virus antigen proteins GD and FD to the nanomagnetic beads containing 10ul binding buffer, mix vertically overnight at room temperature; use a magnet to adsorb the nanomagnetic Beads, remove the supernatant, add 100ul washing buffer II, pipette evenly, the magnetic beads adsorb the nanomagnetic beads, remove the supernatant and wash twice; add 90ul binding buffer, 10ul blocking solution, mix vertically, and react at room temperature for 60 Minutes; Adsorb nano magnetic beads with a magnet, remove the supernatant, add 100ul washing buffer II, and wash once; add 30ul binding buffer, pipette and mix well, set aside;

[0013] 2. Separation of human peripheral blood mononuclear cells: Dilute the collected fresh anticoagulant blood with normal saline 1:1, slowly add it to an equal vol...

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Abstract

The invention relates to a method for screening respiratory syncytial virus memory B cells by a nano magnetic bead process. By adopting the specific antigen-antibody combination theory, respiratory syncytial virus antigen proteins GD and FD are coupled with nano magnetic beads, and separated human peripheral blood lymphocytes are added into the prepared nano-magnetic-bead-containing tube; and since the memory B cells for respiratory syncytial virus in the human peripheralbloodlymphocytes have IgM and IgD on the surface, the respiratory syncytial virus antigen proteins GD and FD coupled with the nano magnetic beads can be specifically combined with the IgM and IgD, thereby separating the memory B cells for respiratory syncytial virus from the human peripheralbloodlymphocytes. The method for screening respiratory syncytial virus memory B cells by nano magnetic beads is simple to operate, has the advantages of high separation speed, high efficiency and high sensitivity, is suitable for quick separation of memory B cells for any virus, and can also be used for comprehensively monitoring RSV (respiratory syncytial virus) infected persons.

Description

Technical field [0001] The invention relates to a screening method for the separation of memory B cells by nano magnetic beads labeled with respiratory syncytial virus antigen. Background technique [0002] Respiratory syncytial virus (RSV) belongs to the genus Paramyxoviridae and is the most common pathogen of lower respiratory tract infections in infants and young children in winter and spring, especially infants from 2 to 6 months old, adults with immunodeficiency and the elderly Humans are also susceptible to RSV. Severe bronchiolitis and pneumonia often occur after RSV infection. Every year, 1‰ to 5‰ of children are hospitalized for RSV worldwide, and the mortality rate of patients is 1‰ to 3‰. Therefore, it is imperative to quickly and comprehensively diagnose and efficiently treat people infected with RSV. [0003] At present, the improvement of rapid detection technology for RSV infection and the research of RSV vaccine and antibodies have become hotspots. The traditional ...

Claims

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Application Information

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IPC IPC(8): C12N5/0781
Inventor 辛洪波陈廷涛邓侃魏强胡萍
Owner NANCHANG UNIV
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