Resumen de Production and utilization of polyclonal antibodies against nisin in an ELISA and for immuno-location of nisin in producing and sensitive bacterial strains
M. Bouksaim, Christophe C. Lacroix, R. Bazin, R. E. Simard
Specific nisin polyclonal antibodies (PAb) were produced in rabbits using nisin Z produced by Lactococcus lactis subsp. lactis biovar diacetylactis UL 719. Antisera were obtained from white female New Zealand rabbits that were first immunized with a nisin Z‐keyhole limpet haemocyanin conjugate and boosted with free nisin Z. Nisin‐specific PAb were purified by affinity chromatography with a yield of 15 mg specific antinisin 100 ml−1 serum. The detection limit of the ELISA test for nisin Z was 0·75 ng ml−1 in buffer but was 1·7 and 3·5 ng ml−1 in milk and complex media broth spiked (5, 10, 20 μg ml−1) with nisin Z, respectively. In nisin Z‐spiked samples, the average concentration was between 90 and 107% of actual added amount. In contrast, when the bioassay (microtitration method) was used, only 50–63% of nisin Z biological activity could be detected. In addition, the affinity‐purified nisin PAb, antirabbit IgG gold conjugate and transmission electron microscopy were successfully used to locate nisin Z on producing cells and to observe its bactericidal effects against sensitive cells.
