PURPOSE: The species cross-reactivity of the monoclonal antibodies infliximab, bevacizumab, and an anti-VEGF-B antibody, 2H10, in humans and rodents was determined. METHODS: The binding of infliximab to human, mouse, and rat TNF-α, of bevacizumab to human, mouse, and rat VEGF-A, and of the 2H10 antibody to human, mouse, and rat VEGF-B was evaluated by ELISA. The sequence of human, mouse, and rat TNF-α and VEGF-A at the binding sites for infliximab and bevacizumab were compared. RESULTS: Infliximab bound to human TNF-α, but no binding to mouse or rat TNF-α was detected between 10 pg/mL and 10 μg/ml. Sequence comparison of the binding site revealed four changes in mouse and five in rat TNF-α compared with human. Bevacizumab bound strongly to human VEGF-A, but showed 5-log weaker binding to both mouse and rat VEGF-A. There was a single amino acid substitution in mouse and rat VEGF-A at the bevacizumab binding site. The 2H10 antibody displayed a similar binding profile to human, mouse, and rat VEGF-B. CONCLUSIONS: The species cross-reactivity of monoclonal antibodies should be determined prior to their use in preclinical animal models. The 2H10 antibody binds to human, mouse, and rat VEGF-B making it suitable for testing in rodent models of human disease.
PURPOSE: The species cross-reactivity of the monoclonal antibodies infliximab, bevacizumab, and an anti-VEGF-B antibody, 2H10, in humans and rodents was determined. METHODS: The binding of infliximab to human, mouse, and rat TNF-α, of bevacizumab to human, mouse, and ratVEGF-A, and of the 2H10 antibody to human, mouse, and ratVEGF-B was evaluated by ELISA. The sequence of human, mouse, and rat TNF-α and VEGF-A at the binding sites for infliximab and bevacizumab were compared. RESULTS:Infliximab bound to human TNF-α, but no binding to mouse or rat TNF-α was detected between 10 pg/mL and 10 μg/ml. Sequence comparison of the binding site revealed four changes in mouse and five in rat TNF-α compared with human. Bevacizumab bound strongly to humanVEGF-A, but showed 5-log weaker binding to both mouse and ratVEGF-A. There was a single amino acid substitution in mouse and ratVEGF-A at the bevacizumab binding site. The 2H10 antibody displayed a similar binding profile to human, mouse, and ratVEGF-B. CONCLUSIONS: The species cross-reactivity of monoclonal antibodies should be determined prior to their use in preclinical animal models. The 2H10 antibody binds to human, mouse, and ratVEGF-B making it suitable for testing in rodent models of human disease.
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Authors: Danielle McAnally; Khandaker Siddiquee; Ahmed Gomaa; Andras Szabo; Stefan Vasile; Patrick R Maloney; Daniela B Divlianska; Satyamaheshwar Peddibhotla; Camilo J Morfa; Paul Hershberger; Rebecca Falter; Robert Williamson; David B Terry; Rafal Farjo; Anthony B Pinkerton; Xiaping Qi; Judith Quigley; Michael E Boulton; Maria B Grant; Layton H Smith Journal: PLoS One Date: 2018-09-12 Impact factor: 3.240
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