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Magnetic microsphere-based methods to study the interaction of teicoplanin with peptides and bacteria

Piyasena, Menake E. and Real, Lilian J. and Diamond, Rochelle A. and Xu, H. Howard and Gomez, Frank A. (2008) Magnetic microsphere-based methods to study the interaction of teicoplanin with peptides and bacteria. Analytical and Bioanalytical Chemistry, 392 (5). pp. 877-886. ISSN 1618-2642. doi:10.1007/s00216-008-2327-y.

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Teicoplanin (teic) from Actinoplanes teichomyceticus is a glycopeptide antibiotic used to treat many Gram-positive bacterial infections. Glycopeptide antibiotics inhibit bacterial growth by binding to carboxy-terminal D-Ala-D-Ala intermediates in the peptidoglycan of the 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 constant for the binding of carboxyfluorescein-labeled D-Ala-D-Ala-D-Ala to teic on microspheres was established via fluorimetry and flow cytometry and was determined to be 0.5x10^-6 and 3.0x10^-6 mol L^-1, respectively. The feasibility of utilizing microparticles with fluorescence methods to detect low levels (the limit of bacterial detection was determined to be 30 colon-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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Additional Information:© Springer-Verlag 2008. Received: 2 June 2008 / Revised: 24 July 2008 / Accepted: 29 July 2008 / Published online: 21 August 2008. The authors gratefully acknowledge financial support for this research by grants from the National Science Foundation (CHE-0515363, CBET-0723271, DMR-0351848 and DMR-0520565), and the National Institutes of Health (1R15AI65468–01 to F.A.G.; P20MD001824 to H.H.X.).
Funding AgencyGrant Number
National Science FoundationCHE-0515363
National Science FoundationCBET-0723271
National Science FoundationDMR-0351848
National Science FoundationDMR-0520565
National Institutes of Health1R15AI65468–01
National Institutes of HealthP20MD001824
Subject Keywords:bioanalytical methods; magnetic microspheres; teicoplanin; fluorescence; microfluidics
Issue or Number:5
Record Number:CaltechAUTHORS:PIYabc08
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Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:13386
Deposited By: Tony Diaz
Deposited On:04 May 2009 16:45
Last Modified:08 Nov 2021 22:37

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