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Clinical and Diagnostic Laboratory Immunology, January 2005, p. 93-100, Vol. 12, No. 1
1071-412X/05/$08.00+0     doi:10.1128/CDLI.12.1.93-100.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Inhibition of Biofilm Formation by Monoclonal Antibodies against Staphylococcus epidermidis RP62A Accumulation-Associated Protein

Department of Chemical Engineering, University of Connecticut, Storrs,¹ Center of Biomaterials, University of Connecticut Health Center, Farmington, Connecticut,³ Hybridoma/Phage Display Core Facility, University of Alabama-Huntsville, Huntsville, Alabama²

Received 28 May 2004/ Returned for modification 21 July 2004/ Accepted 12 August 2004

Staphylococcus epidermidis expresses a 140-kDa cell wall-bound protein accumulation-associated protein (AAP) to adhere to and accumulate as a biofilm on a surface. Potentially blocking AAP with a monoclonal antibody (MAb) could reduce or eliminate S. epidermidis bacterial colonization of biomedical devices. Here, we report on our efforts to (i) isolate AAP, (ii) generate MAbs against AAP, and (iii) determine the efficacy of MAbs to inhibit S. epidermidis biofilm formation. An M7 S. epidermidis mutant, reportedly deficient in AAP expression, was used as a negative control. Postinoculation murine sera, containing polyclonal antibodies against AAP, were able to reduce S. epidermidis biofilm formation by 54%. Select MAbs against AAP were able to reduce S. epidermidis by no more than 66%. Two MAb mixtures, 12C6/12A1 and 3C1/12A1, reduced S. epidermidis accumulation up to 79 and 87%, respectively, significantly more than individual MAbs. Contrary to a previous report, biofilm-deficient S. epidermidis mutant M7 expressed a 200-kDa protein on its cell wall that specifically bound AAP MAbs. Peptide characterization of this M7 protein by microcapillary reversed-phase high-pressure liquid chromatography-nanoelectrospray tandem mass spectrometry resulted in 53% homology with AAP. Ongoing studies will elucidate the dynamic expression of AAP and the M7 200-kDa protein in order to define their roles in biofilm formation.