Magnetic microsphere-based methods to study the interaction of teicoplanin with peptides and bacteria.
Magnetic microsphere-based methods to study the interaction of teicoplanin with peptides and bacteria.
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DOI：
10.1007/s00216-008-2327-y
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发表时间：
2008-11
影响因子：
4.3
通讯作者：
Gomez, Frank A.
中科院分区：
文献类型：
--
作者：
Piyasena, Menake E.;Real, Lilian J.;Diamond, Rochelle A.;Xu, H. Howard;Gomez, Frank A.
Teicoplanin （teic）是一种糖肽类抗生素，用于治疗革兰氏阳性细菌感染。糖肽类抗生素通过与革兰氏阳性菌细胞壁肽聚糖中羧基末端的D-Ala-D-Ala中间体结合抑制细菌生长。本文报道了磁性微球与teic （teic-microspheres）的衍生化。基于荧光的技术已经被开发用来分析微球与两个D-Ala-D-Ala端肽的结合特性。羧基荧光素标记的D-Ala-D-Ala-D-Ala在微球上与teic结合的解离常数分别为0.5 × 10 - 6和3.0 × 10 - 6 M。利用微粒与荧光方法检测低水平（细菌检测限被确定为200菌落形成单位[cfu]）革兰氏阳性细菌的可行性已得到证实。一个简单的微流体实验证明了开发基于微球的亲和分析来研究肽-抗生素相互作用的可能性。
Teicoplanin (teic) from Actinoplanes teichomyceticus is a glycopeptide antibiotic used to treat many gram-positive bacterial infections. Glycopeptide antibiotics inhibit the bacteria growth by binding to carboxy-terminal D-Ala-D-Ala intermediates in the peptidoglycan of cell wall of gram positive bacteria. In this paper we report the derivatization of magnetic microspheres with teic (teic-microspheres). Fluorescence based techniques have been developed to analyze the binding properties of the microspheres to two D-Ala-D-Ala terminus peptides. The dissociation constants for the binding of carboxyfluorescein labeled D-Ala-D-Ala-D-Ala to teic on microspheres is established via fluorometry and flow cytometry and was determined to be 0.5 × 10−6 and 3.0 × 10−6 M, respectively. Feasibility of utilizing microparticles with fluorescence methods to detect low levels (limit of bacterial detection was determined to be 200 colony forming units [cfu]) of gram-positive bacteria has been demonstrated. A simple microfluidic experiment is reported to demonstrate the possibility of developing microsphere based affinity assays to study peptide-antibiotic interaction.
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