Production of polyclonal and monoclonal antibodies against gill-associated virus and the development of an ELISA

Abstract

Gill-associated virus (GAV) is the type species of the genus Okavirus in the Roniviridae of the Nidovirales and has been associated with mortalities in farmed Penaeus monodon (black tiger prawn). Polyclonal antibodies (PAbs) and monoclonal antibodies (MAbs) specific to GAV were produced following immunisation with protein subunits of purified GAV isolated from the gill tissues of P. monodon into either a six week old chicken (PAbs) or into a six week old Balb/c mouse for the production of the MAbs. Western blot analysis indicated that the PAbs reacted with 63, 110 and 170 kDa proteins, while the MAbs reacted to the 20 kDa protein of GAV. The MAbs were further investigated to ensure specificity by immunohistochemistry and elution of affinity purified GAV protein from the MAbs. An enzyme linked immunosorbent assay was then developed using the prepared PAbs and MAbs for the detection of GAV.