Author: DOKI, Tomoyoshi; TAKANO, Tomomi; HOHDATSU, Tsutomu
Title: Development of a mouse-feline chimeric antibody against feline tumor necrosis factor-alpha Document date: 2016_6_4
ID: y0x0quha_16
Snippet: The V H (414 bp) and V L (378 bp) genes were amplified from cDNA derived from hybridoma mAb 2-4 (Fig. S2) , and the C H gene (1005 bp) was amplified from cDNA derived from feline PBMCs. The C L gene (330 bp) was amplified using a plasmid containing a known C L gene as a template. Each PCR product was cloned into the pCR-blunt II-TOPO vector and sequenced. The V H and C H genes were ligated, inserted into the pCDNA 3.1 (+)/Neo expression vector an.....
Document: The V H (414 bp) and V L (378 bp) genes were amplified from cDNA derived from hybridoma mAb 2-4 (Fig. S2) , and the C H gene (1005 bp) was amplified from cDNA derived from feline PBMCs. The C L gene (330 bp) was amplified using a plasmid containing a known C L gene as a template. Each PCR product was cloned into the pCR-blunt II-TOPO vector and sequenced. The V H and C H genes were ligated, inserted into the pCDNA 3.1 (+)/Neo expression vector and sequenced (Fig. S3) . The amino acid sequence deduced from the base sequence was confirmed to show the characteristics of the variable region of mAb 2-4 and the constant region of the feline immunoglobulin heavy chain. Similarly, the amino acid sequences of the V L and C L genes were deduced from the base sequences, and it was confirmed that the recombinant protein had the characteristics of the variable region of mAb 2-4 and the constant region of the feline immunoglobulin kappa light chain (Fig. S4) .
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