Synthetic Peptide Coupled to KLH Elicits Antibodies Against β8 Integrin

The β8 integrin, which forms αvβ8 heterodimers, is being widely investigated because of its specific structure and functions compared with other integrins. In this report, a 12 aa-long peptide of β8 integrin cytoplasmic domain was synthesized according to a published sequence and covalently coupled to keyhole limpet hemocyanin (KLH). Three stable strains of hybridomas (3G6, 5C7, 5H3) that can secrete high specific monoclonal antibodies against β8 integrin were successfully established by hybridoma technique. The isotypes of these MAbs were tested to be IgG2a. Their characterizations were shown by enzyme-linked immunosorbent assay (ELISA), Western blot analysis, and immunocytochemistry (ICC). The affinity constants (Kaff ) of the MAbs 3G6, 5C7, and 5H3 were measured by non-competitive ELISA respectively. Western blot analyses and immunocytochemistry demonstrated that all the MAbs were directed against β8 integrin with high specificity.