Method for establishing HIV (human immunodeficiency virus) virus antibody yeast display library, method for screening virus broad-spectrum neutral antibody and application thereof

Abstract

The invention provides a method for screening a virus broad-spectrum neutral antibody and application thereof. The method comprises the following steps: preparing or providing a virus antigen protein; marking the virus antigen protein by using a fluorescence marker; preparing a cDNA (complementary deoxyribonucleic acid) template of a virus positive patient, designing primers, and amplifying the antibody gene; establishing a yeast antibody display library by using the obtained antibody gene; carrying out antigen protein mixed incubation on yeast cells in the obtained antibody display library and the obtained virus with the fluorescence marker, setting a cell separation door, and separating fluorescence positive yeast cells by a flow cytometry; and carrying out sequencing identification on the fluorescence positive yeast cells to obtain the virus broad-spectrum neutral antibody. The invention also provides a method for establishing an HIV (human immunodeficiency virus) virus antibody yeast display library and application thereof.

Description

technical field [0001] The invention relates to the field of immunology, in particular to a method for constructing an HIV virus antibody yeast display library, a method for screening virus broad-spectrum neutralizing antibodies and applications thereof. Background technique [0002] Broad-spectrum virus neutralizing antibodies are immunoglobulins acquired by infected patients. This type of antibody broadly binds to virus particles, preventing them from further infecting human cells, and can lead to cleavage of virus particles, thereby causing a "neutralization" response. It is especially important in the treatment of mutating viruses, such as HIV. At present, to screen virus broad-spectrum neutralizing antibodies, patients’ peripheral blood mononuclear lymphocytes (PBMCs) are mostly used to amplify antibody DNA, and phage display is used to perform repeated screening in sequence to establish a phage display antibody library. [0003] However, since the proportion of neutra...

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Problems solved by technology

[0005] In order to solve the above problems, the first aspect of the present invention provides a method for screening virus broad-spectrum neutralizing antibodies, the method is by constructing a yeast antibody display library, using fluorescently labeled viral antigen proteins, and combining flow cytometry sorting technology Screening of virus broad-spectrum neutralizing antibodies, the flow sorting process is a visual screening, by setting the cell separation gate, the controllability of the screening process is improved, and the problem of excessive false positives caused by conventional screening is avoided

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