Production and utilization of polyclonal antibodies against nisin in an ELISA and for immuno-location of nisin in producing and sensitive bacterial strains
| Autor: | M. Bouksaim, R. Bazin, Ronald E. Simard, Christophe Lacroix |
|---|---|
| Rok vydání: | 1999 |
| Předmět: |
Enzyme-Linked Immunosorbent Assay
Biology Applied Microbiology and Biotechnology Chromatography Affinity Microbiology chemistry.chemical_compound Bacteriocins Bacteriocin Affinity chromatography polycyclic compounds Animals Bioassay Nisin Antiserum Immune Sera Lactococcus lactis food and beverages General Medicine biochemical phenomena metabolism and nutrition biology.organism_classification Streptococcaceae Microscopy Electron Milk chemistry Polyclonal antibodies Immunoglobulin G biology.protein bacteria Female lipids (amino acids peptides and proteins) Rabbits Biotechnology |
| Zdroj: | Journal of Applied Microbiology. 87:500-510 |
| ISSN: | 1364-5072 |
| DOI: | 10.1046/j.1365-2672.1999.00842.x |
| Popis: | 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 microg 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. |
| Databáze: | OpenAIRE |
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