Dual specific binding proteins directed against immune cell receptors and tlr signaling autoantigens
A binding protein, bispecific technology, applied in immunoglobulin, anti-animal/human immunoglobulin, anti-receptor/cell surface antigen/cell surface determinant immunoglobulin, etc.
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example 1
[0212] Example 1: Anti-mouse IgM and anti-DNA dual variable domain immunoglobulin (DVD-Ig TM ) protein generation and characterization
[0213] Four-chain dual variable domain immunoglobulin (DVD -Ig TM )protein. DVD-Ig TM The constructs were cloned and expressed in human embryonic kidney 293 cells and purified according to methods known in the art. for DVD-Ig TM The VH and VL chain amino acid sequences of the proteins are provided in Table 1. The SEQ ID NO listed in the leftmost column of Table 2 refers to the DVD-Ig in that row of the table TM The sequence of the complete variable domains of the heavy and light chains of the protein. Each row in the rightmost column of Table 2 provides three SEQ ID NOs. The first number refers to the SEQ ID NO of the outer variable domain sequence, the second number refers to the SEQ ID NO of the linker, and the third number refers to the SEQ ID NO of the inner variable domain sequence, which are common in a complete DVD-Ig TM Varia...
example 2
[0221] Example 2: For the determination of parental antibodies and DVD-Ig TM Determination of functional activity of proteins
example 21
[0222] Example 2.1: Mice
[0223] AM14 B cell receptor (BCR) transgenic mice have been described previously (Sweet et al. (2010) Autoimmun. 43(8):607-18). Briefly, AM14 is a BCR comprising AM14 heavy chain and Vk8 light chain that recognizes the "a" allotype of murine IgG2a (mIgG2a). 95-98% of B cells in mice positive for AM14 heavy chain and Vk8 light chain express AM14BCR. To generate AM14B cells deficient in TLR9, TLR7 or FcyRIIB, AM14 and Vk8 genes were bred to appropriate knockout mice.
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