Yao R C, Mahoney D F
Lilly Research Laboratories, Eli Lilly & Company, Indianapolis, Indiana 46285.
Appl Environ Microbiol. 1989 Jun;55(6):1507-11. doi: 10.1128/aem.55.6.1507-1511.1989.
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.
通过使用在经与钥孔戚血蓝蛋白共价连接的23-氨基-O- mycaminosyltylonolide（23-氨基-OMT）免疫的兔中产生的多克隆抗体，开发了一种用于检测大环内酯类抗生素的酶联免疫吸附测定法。在直接竞争酶联免疫吸附测定中，使用与碱性磷酸酶偶联的23-氨基-OMT作为酶联标记来表征该抗体的特异性和敏感性。该测定法对23-氨基-OMT的灵敏度低至0.3 ng/ml，50%抑制浓度为8 ng/ml。该抗体与具有氨基取代糖部分的12元、14元或16元大环内酯类表现出良好的反应性，无论是否存在中性糖残基。与大环内酯环结构（泰乐内酯）或仅含中性糖的大环内酯类几乎没有交叉反应。与多烯类或非大环内酯类抗生素未观察到交叉反应。在发酵液中生长的已知大环内酯产生菌培养物也表现出良好的反应性，表明该测定法可用于检测发酵中的这类代谢产物。
