Purification of adenovirus 4 type-specific hemagglutinins for use in diagnostic counterelectrophoresis tests

Three type-specific hemagglutinins of adenovirus (AV) 4 were purified, tested for various activities, and used as immunogens in rabbits. The components were purified by anion exchange and exclusion chromatographies to purification factors of 33, 20, and 37, respectively, and were identified as fiber (polypeptide, mol wt 70,000; incomplete hemagglutinin), penton (mol wt greater than 400,000; incomplete hemagglutinin; possession of cytotoxic activity), and dodecon (mol wt greater than 400,000; complete hemagglutinin; appearance by electron microscopy). Rabbit antisera to these hemagglutinating components possessed type-specific hemagglutination-inhibition and serum neutralization antibodies and were thoroughly evaluated for potency and specificty in immunodiffusion (ID), immunoelectrophoresis, and counterelectrophoresis (CE) tests. The anti-fiber and anti-dodecon sera were highly type-specific in ID and CE tests with a battery of commonly isolated AV serotypes; the anti-penton serum was similarly specific after it was absorbed with AV 2 hexons. These sera were thus proved to be suitable reagents for use in both the routine diagnostic ID test and the more rapid CE test.