A novel and convenient method to immunize animals: Inclusion bodies from recombinant bacteria as antigen to directly immunize animals

Abstract

Purified recombinant protein has been routinely used to immunize rabbits to produce polyclonal antibodies. The process of the purification of recombinant proteins from bacterial inclusion bodies is both labor intensive and time consuming. To determine whether whole inclusion bodies can be used as antigen in polyclonal antibody production, we amplified Gallus gallus phospholipase A2 gene by RTPCR, fused with glutathione-S-transferase (GST), and expressed recombinant Escherichia coli strain Rosseta (DE3) in the form of inclusion bodies. The inclusion bodies isolated from the bacteria cells were directly injected into rabbits and hens at 300 μg dose four times. Western blot was performed to detect antibodies in serum and in egg yolk, which revealed strong immunoreactions. The results suggested that using recombinant protein-containing inclusion bodies as antigen is a novel and convenient method to immunize animals for polyclonal antibody production.

Key words: Polyclonal antibodies, inclusion bodies, antigen, recombinant phospholipase A2.