High affinity neutralizing monoclonal antibodies to programmed death ligand 1 (pd-l1) and uses thereof
A PD-L1, antibody technology, applied in the direction of antibody, anti-receptor/cell surface antigen/cell surface determinant immunoglobulin, anti-tumor drugs, etc., can solve the problem of reaching the effective level of chemotherapeutic agents, but the results are not satisfactory People's Choice, etc.
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[0148] Transgenic mice overexpressing mouse Ig-α, mouse Ig-β, and human interleukin-6 were injected intraperitoneally with recombinant human PD-L1 (R&D Systems) at 2-week intervals. After establishing a significant immune response as measured by serum ELISA, lymph node, spleen, and bone marrow cells were harvested, B cells expressing IgM isotype antibodies on the surface were subtracted using magnetic beads, and the remaining cells were targeted using a MoFlo fluorescence-activated cell sorter. Cells were sorted for their ability to bind PD-L1.
[0149] Cells positive for PD-L1 binding were sorted into 96-well plates, single-cell RT-PCR was performed to amplify the variable region, and the variable region was cloned into a heavy chain human IgG1 or IgG4 constant region or light chain In the expression vector of human Igk constant region. The resulting heavy and light chain clonal pairs were transfected into CHO cells, and the resulting antibody proteins were purified using pr...
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