Production and purification of polyclonal antibody against shrimp lectin LvLTLC1

Lectins play an important role in the shrimp immune response owing to the ability of recognising and eliminating invasive microorganisms. LvLTLC1, a recently discovered lectin, is able to resist the effects of Vibrio parahaemolyticus causing Acute Hepatopancreatic Necrosis Disease (AHPND) by its potential bacteria clearance of Vibrio spp. in vivo. In this research, the preparation of LvLTLC1 antigen by expressing and purifying the His-tagged recombinant protein, and production of LvLTLC1-specific mouse polyclonal antibodies were conducted. The produced mouse polyclonal antibodies were then used to determine the antiserum or purified antibodies titer by indirect ELISA and detect the LvLTLC1 expression in hepatopancreas tissues of both AHPND-infected and uninfected shrimps by SDS-PAGE and Western blot analysis, respectively. Results showed that LvLTLC1 expression in hepatopancreas tissues of AHPND-infected shrimps was significantly higher than that of healthy shrimps. Collectively, we reported for the first time the generation and evaluation of LvLTLC1-specific mouse antibody as a tool for further studies on understanding the nature of immune response in shrimp and developing immune-based applications.