Abstract
Monoclonal antibodies were prepared against nisin A (nisA), an antimicrobial peptide that is used as a food preservative and is produced by strains of Lactococcus lactis subsp. lactis. NisA antibodies were secreted by hybridomas derived from mice immunized with a nisA−keyhole limpet hemocyanin conjugate. Competitive direct enzyme-linked immunoabsorbent assays (CD-ELISA) were devised whereby free nisA and nisA−horseradish peroxidase competed for antibody binding. The detection limit for nisA in this assay was 10 ng/mL. Antibodies also cross-reacted with nisin Z (nisZ). Mean concentrations of nisA and nisZ required to inhibit 50% antibody binding for the selected clones were 1180 and 3750 ng/mL, respectively. When the nisA CD-ELISA performance was assessed in spiked (250, 1250, and 5000 ng/g) spread cheese, the average recovery was 116.5%, with mean intra- and interassay coefficients of variation of 13.3 and 14.1%, respectively. Keywords: Nisin A; monoclonal antibodies; ELISA; lantibiotics; bacteriocins
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