Abstract
An enzyme-linked immunosorbent assay was developed for the detection of macrolide antibiotics by using a polyclonal antibody generated in rabbits immunized with 23-amino-O-mycaminosyltylonolide (23-amino-OMT) covalently linked to keyhole limpet hemocyanin. The specificity and sensitivity of this antibody were characterized by using 23-amino-OMT coupled to alkaline phosphatase as an enzyme-linked label in a direct competitive enzyme-linked immunosorbent assay. The assay sensitivity was as low as 0.3 ng/ml for 23-amino-OMT, with a 50% inhibitory concentration of 8 ng/ml. This antibody exhibited good reactivity with 12-, 14- or 16-membered macrolides possessing amino-substituted sugar moieties, regardless of the presence of neutral sugar residues. Little or no cross-reactivity was observed with the macrocyclic lactone ring structure (tylactone) or macrolides containing only neutral sugars. No cross-reaction was observed with polyenes or nonmacrolide antibiotics. Known macrolide-producing cultures grown in fermentation broth also showed good reactivity, indicating that this assay is useful in detecting this class of metabolites in fermentation.
Full text
PDF
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Avrameas S. Coupling of enzymes to proteins with glutaraldehyde. Use of the conjugates for the detection of antigens and antibodies. Immunochemistry. 1969 Jan;6(1):43–52. doi: 10.1016/0019-2791(69)90177-3. [DOI] [PubMed] [Google Scholar]
- Baltz R. H., Seno E. T. Properties of Streptomyces fradiae mutants blocked in biosynthesis of the macrolide antibiotic tylosin. Antimicrob Agents Chemother. 1981 Aug;20(2):214–225. doi: 10.1128/aac.20.2.214. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hebert G. A., Pelham P. L., Pittman B. Determination of the optimal ammonium sulfate concentration for the fractionation of rabbit, sheep, horse, and goat antisera. Appl Microbiol. 1973 Jan;25(1):26–36. doi: 10.1128/am.25.1.26-36.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kirst H. A., Toth J. E., Wind J. A., Debono M., Willard K. E., Molloy R. M., Paschal J. W., Ott J. L., Felty-Duckworth A. M., Counter F. T. Synthesis, antimicrobial evaluation and structure-activity relationships within 23-modified derivatives of 5-O-mycaminosyltylonolide. J Antibiot (Tokyo) 1987 Jun;40(6):823–842. doi: 10.7164/antibiotics.40.823. [DOI] [PubMed] [Google Scholar]
- Shimada N., Ueda T., Yokoshima T., Umemura K., Shomura T. Radioimmunoassay for metabolites of 9,3"-diacetylmidecamycin, a macrolide antibiotic. Antimicrob Agents Chemother. 1982 Dec;22(6):976–980. doi: 10.1128/aac.22.6.976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tanaka Y., Kimura K., Komagata Y., Tsuzuki K., Tomoda H., Omura S. Radioimmunoassay for erythromycin derivatives. J Antibiot (Tokyo) 1988 Feb;41(2):258–260. doi: 10.7164/antibiotics.41.258. [DOI] [PubMed] [Google Scholar]
- Voller A., Bidwell D. E., Bartlett A. Enzyme immunoassays in diagnostic medicine. Theory and practice. Bull World Health Organ. 1976;53(1):55–65. [PMC free article] [PubMed] [Google Scholar]
- Yao R. C., Mahoney D. F. Enzyme-linked immunosorbent assay for the detection of fermentation metabolites: aminoglycoside antibiotics. J Antibiot (Tokyo) 1984 Nov;37(11):1462–1468. doi: 10.7164/antibiotics.37.1462. [DOI] [PubMed] [Google Scholar]