Production of antibodies against enterocin B for immunological detection and purification purposes

Polyclonal antibodies were generated against the bacteriocin enterocin B by conjugating enterocin B (ENT) to bovine serum albumin (BSA) through a bifu nctional crosslinking agent. Rabbits immunized with the ENT-BSA conjugates developed antibodies specific for enterocin B. Non-competive indirect (NCI) ELISAs were able to detect the presence of enterocin B in the supernatant of a producer strain and from a partially-purified sample. Competitive indi rect (CI) ELISAs demonstrated concentrations of enterocin B ranging from 40 0-1600 ng ml(-1) were required for 50% antibody binding inhibition. An immu noaffinity chromatography column was developed by linking the antibodies to CNBr-activated Sepahrose 4B. The purification method yielded pure enteroci n; however, the harsh elution conditions did not render a regeneratable col umn.