Neutralizing antibody for resisting influenza A virus and application thereof
A technology of influenza A virus and influenza A, applied in the direction of antiviral agent, virus/bacteriophage, antiviral immunoglobulin, etc., can solve the problems of not neutralizing or inhibiting influenza A virus infection or remission, and not yet discovered
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[0102] Antibody preparation
[0103] Antibodies according to the invention can be prepared by any method known in the art. Common methods for the production of monoclonal antibodies, eg using hybridoma technology, are known (Kohler, G. and Milstein, C, 1975; Kozbar et al. 1983). In one example, the alternative EBV immortalization method described in WO2004 / 076677 is used.
[0104] B cells producing antibodies of the invention can be transformed with EBV in the presence of polyclonal B cell activators using the methods described in WO2004 / 076677. Transformation with EBV is a standard technique and can be easily adapted to include polyclonal B cell activators.
[0105] Optionally, additional stimulators of cell growth and cell differentiation can be added during the transformation step to further increase efficiency. These stimuli can be cytokines such as IL-2 and IL-15. In one aspect, IL-2 is added during the immortalization step to further improve the efficiency of immor...
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[0188] Exemplary embodiments of the invention are provided in the following examples. The following examples are given by way of illustration only and to assist those of ordinary skill in using the invention. The examples are not intended to otherwise limit the scope of the invention in any way.
example 1
[0189] Example 1: Generation and Characterization of Influenza A Broadly Neutralizing Antibodies from Plasma Cells
[0190] To identify individuals who can develop heterosubtypic antibodies in response to seasonal influenza vaccines (containing H1 and H3 HA), we directed circulating plasma cells collected on day 7 after boost to secrete conjugate vaccine or unrelated H5 The ability of the antibody to HA (A / VN / 1203 / 04) was screened by ELISPOT. Surprisingly, however, no H5-specific plasma cells were detected in 4 of 5 donors tested, and 14% of IgG-secreting plasma cells in one donor produced antibodies against H5, And 57 percent developed antibodies against the vaccine. CD138+ plasma cells were isolated from peripheral blood mononuclear cells (PBMC) collected on day 7 post-vaccination by using magnetic microbeads followed by cell sorting using a FACSAria instrument. A limited number of plasma cells are seeded into microwell culture plates. Culture supernatants were tested i...
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