Production of Antibodies against Enterocin B for Immunological Detection and Purification Purposes

Abstract

Polyclonal antibodies were generated against the bacteriocin enterocin B by conjugating enterocin B (ENT) to bovine serum albumin (BSA) through a bifunctional 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 indirect (CI) ELISAs demonstrated concentrations of enterocin B ranging from 400-1600 ng ml⁻¹ were required for 50% antibody binding inhibition. An immunoaffinity chromatography column was developed by linking the antibodies to CNBr-activated Sepahrose 4B. The purification method yielded pure enterocin; however, the harsh elution conditions did not render a regeneratable column.

Keywords:

• Crosslinking Agent
• Bacteriocins
• Purification
• Detection
• Enzyme Immunoassay