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Next Article 

Clinical and Vaccine Immunology, August 2009, p. 1095-1104, Vol. 16, No. 8
1071-412X/09/$08.00+0     doi:10.1128/CVI.00085-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Selection and Characterization of Murine Monoclonal Antibodies to Staphylococcus aureus Iron-Regulated Surface Determinant B with Functional Activity In Vitro and In Vivo{triangledown} ,{dagger}

Martha Brown, Rose Kowalski, Julie Zorman, Xin-min Wang, Victoria Towne, Qinjian Zhao, Susan Secore, Adam C. Finnefrock, Tim Ebert, Greg Pancari, Kevin Isett, Yuhua Zhang, Annaliesa S. Anderson,{ddagger} Donna Montgomery, Leslie Cope, and Tessie McNeely*

Merck Research Labs, Merck and Co., Inc., 770 Sumneytown Pike, West Point, Pennsylvania 19486

Received 27 February 2009/ Returned for modification 23 March 2009/ Accepted 12 June 2009

In an effort to characterize important epitopes of Staphylococcus aureus iron-regulated surface determinant B (IsdB), murine IsdB-specific monoclonal antibodies (MAbs) were isolated and characterized. A panel of 12 MAbs was isolated. All 12 MAbs recognized IsdB in enzyme-linked immunosorbent assays and Western blots; 10 recognized native IsdB expressed by S. aureus. The antigen epitope binding of eight of the MAbs was examined further. Three methods were used to assess binding diversity: MAb binding to IsdB muteins, pairwise binding to recombinant IsdB, and pairwise binding to IsdB-expressing bacteria. Data from these analyses indicated that MAbs could be grouped based on distinct or nonoverlapping epitope recognition. Also, MAb binding to recombinant IsdB required a significant portion of intact antigen, implying conformational epitope recognition. Four MAbs with nonoverlapping epitopes were evaluated for in vitro opsonophagocytic killing (OPK) activity and efficacy in murine challenge models. These were isotype switched from immunoglobulin G1 (IgG1) to IgG2b to potentially enhance activity; however, this isotype switch did not appear to enhance functional activity. MAb 2H2 exhibited OPK activity (≥50% killing in the in vitro OPK assay) and was protective in two lethal challenge models and a sublethal indwelling catheter model. MAb 13C7 did not exhibit OPK (<50% killing in the in vitro assay) and was protective in one lethal challenge model. Neither MAb 13G11 nor MAb 1G3 exhibited OPK activity in vitro or was active in a lethal challenge model. The data suggest that several nonoverlapping epitopes are recognized by the IsdB-specific MAbs, but not all of these epitopes induce protective antibodies.

* Corresponding author. Mailing address: Merck and Co., Inc., WP26-253, West Point, PA 19486. Phone: (215) 652-6752. Fax: (215) 993-1540. E-mail: tessie_mcneely{at}merck.com

{triangledown} Published ahead of print on 24 June 2009.

{dagger} Supplemental material for this article may be found at http://cvi.asm.org/.

{ddagger} Present address: Wyeth Research, 401 N. Middletown Road 205/3112, Pearl River, NY 10965.

Clinical and Vaccine Immunology, August 2009, p. 1095-1104, Vol. 16, No. 8
1071-412X/09/$08.00+0     doi:10.1128/CVI.00085-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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