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Journal of Virology, November 2001, p. 10906-10911, Vol. 75, No. 22
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.22.10906-10911.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Fine Definition of the Epitope on the gp41 Glycoprotein of Human Immunodeficiency Virus Type 1 for the Neutralizing Monoclonal Antibody 2F5

Carol E. Parker,1,* Leesa J. Deterding,1,* Christine Hager-Braun,1 James M. Binley,2 Norbert Schülke,3 Hermann Katinger,4 John P. Moore,2 and Kenneth B. Tomer1

Laboratory of Structural Biology, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, North Carolina 277091; Department of Microbiology and Immunology, Joan and Sanford I. Weill Medical College of Cornell University, New York, New York 100212; Progenics Pharmaceuticals Inc., Tarrytown, New York 105913; and Institute for Applied Microbiology, University of Agriculture and Forestry, 1190 Vienna, Austria4

Received 9 May 2001/Accepted 13 August 2001

Matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS), in combination with proteolytic protection assays, has been used to identify the functional epitope on human immunodeficiency virus envelope glycoprotein gp41 for the broadly neutralizing anti-gp41 human monoclonal antibody 2F5. In this protection assay-based procedure, a soluble gp140 protein with a stabilizing intermolecular disulfide bond between the gp120 and gp41 subunits (SOS gp140) was affinity bound to immobilized 2F5 under physiological conditions. A combination of proteolytic enzymatic cleavages was then performed to remove unprotected residues. Residues of SOS gp140 protected by their binding to 2F5 were then identified based on their molecular weights as determined by direct MALDI-MS of the immobilized antibody beads. The epitope, NEQELLELDKWASLWN, determined by this MALDI-MS protection assay approach consists of 16 amino acid residues near the C terminus of gp41. It is significantly longer than the ELDKWA core epitope previously determined for 2F5 by peptide enzyme-linked immunosorbent assay. This new knowledge of the structure of the 2F5 epitope may facilitate the design of vaccine antigens intended to induce antibodies with the breadth and potency of action of the 2F5 monoclonal antibody.

* Corresponding author. Mailing address for Leesa J. Deterding: Laboratory of Structural Biology, National Institute of Environmental Health Sciences, National Institutes of Health, P.O. Box 12233, Research Triangle Park, NC 27709. Phone: (919) 541-3009. Fax: (919) 541-0220. E-mail: deterding{at}niehs.nih.gov. Present address for Carol E. Parker: Dept. of Biochemistry and Biophysics, CB7260, University of North Carolina School of Medicine, Chapel Hill, NC 27599. Phone: (919) 966-9989. Fax: (919) 966-2852. E-mail: Carol_Parker{at}med.unc.edu.

Journal of Virology, November 2001, p. 10906-10911, Vol. 75, No. 22
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.22.10906-10911.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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Copyright © 2001 by the American Society for Microbiology. All rights reserved.