Polyclonal antibody generation in rabbit by administration of an Organophosphorus Acid Anhydrolase (OPAA) from squid

Abstract

When a nerve gas hydrolyzing enzyme [organophosphorus acid anhydrolase (OPAA), formerly DFPase] purified from squid hepatopancreas was injected into rabbits, the resulting sera (RAS) inhibited OPAA purified from either squid hepatopancreas or squid optic ganglia. The inhibition was non-competitive, with 50% inhibition at a 1:1,000 serum dilution, and with the limit of inhibition (in effect, a “titer”) at approximately 1:10,000. This RAS did not inhibit the distinctly different OPAAs from a mammalian and two bacterial sources. The hepatopancreas-generated RAS also reacted positively to the appropriate enzyme-linked immunosorbent assay (ELISA) at a titer of 1:100,000. In marked contrast, when OPAA purified from squid optic ganglion was injected into rabbits, the resulting sera did not inhibit squid OPAA, and did not give a positive ELISA. Control sera taken from the same rabbits prior to any injection (RS) did not inhibit the OPAAs. These results show another major difference between squid type OPAAs and the OPAAs from other sources, sometimes termed “Mazur type” OPAAs.