Synthetic Peptide Coupled to KLH Elicits Antibodies Against β8 Integrin

The beta 8 integrin, which forms alpha v beta 8 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 beta 8 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 beta 8 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 beta 8 integrin with high specificity.