Neutralizing antibodies against SARS-CoV-2 without ADE effect
A sars-cov-2, antibody technology, applied in the direction of antibodies, applications, antiviral agents, etc.
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[0336] Preparation of immunogenic antigens and production of monoclonal antibodies can be performed by any suitable technique, such as recombinant protein production. Immunogenic antigens can be administered to animals in the form of purified proteins or protein mixtures, including whole cells, cell extracts or tissue extracts, or antigens can be formed de novo in animals from nucleic acids encoding the antigens or portions thereof.
[0337] Another embodiment of the invention is an isolated polynucleotide encoding any one of the antibody heavy chain variable regions and / or the antibody light chain variable regions of the invention. Multiple polynucleotides encoding the same antibody of the invention due to genetic code degeneracy or codon preference in a given expression system are also within the scope of the invention. A polynucleotide sequence encoding the VH or VL of an antibody of the invention, or fragments thereof, may be operably linked to one or more regulatory eleme...
Embodiment 1
[0400] Example 1: Serological response to SARS-CoV-2
[0401] 16 donors who had recovered from SARS-CoV-2 infection and 8 healthy donors recruited before the outbreak of COVID-19 ( figure 2 ) to collect serum samples. Sera from post-infection recovered donors showed significantly higher ELISA binding potencies to individual SARS-CoV-2 S-protein domains (RBD, S1, S2, and S-ECD) compared to unexposed healthy donors ( image 3 A-3D; figure 1 ). After infection, the N-protein in the virion also elicits a robust antibody response ( image 3 E). To determine neutralizing activity in convalescent sera, their ability to block viral infection was tested in Huh-7 cells by luciferase-expressing SARS-CoV-1 or SARS-CoV-2 pseudoviruses (Xia et al. , 2020a; Xia et al., 2020b). The luciferase signal (a surrogate for infection) was then compared at different dilutions of serum or purified IgG antibody ( image 3 F-3H). Although 5 individuals (donors #5, #6, #7, #8 and #16) achieved...
Embodiment 2
[0403] Example 2: Human monoclonal antibodies against SARS-CoV-2 S-protein
[0404] To characterize the IgG antibodies responsible for potent neutralizing activity in this selected individual, SARS-CoV-2 RBD- or S-ECD-binding B cells were identified using a dual fluorescent dye labeling strategy ( Figure 4 ) (Wang, Q. et al., 2020). Unexposed uninfected controls showed background levels of bait protein-specific B cells, whereas donor #16 with high serum neutralizing activity showed a different population of bait protein-binding B cells. Avi-Tag-tagged biotinylated RBD, chemically biotinylated S-ECD, and Avi-Tag-tagged biotinylated S-ECD bait protein stained B cells at frequencies of 0.025%, 0.12%, and 0.21%, respectively, as background stained about 6 to 20 times ( Figure 5 A-5C).
[0405] Gated double positive cells (bait protein-PE + and bait protein-APC + ) by single cell sorting, and immunoglobulin heavy chain ( IGH ; IgG isotype) and light chain ( IGL or IG...
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