Journal
JOURNAL OF VIROLOGY
Volume 75, Issue 22, Pages 10906-10911Publisher
AMER SOC MICROBIOLOGY
DOI: 10.1128/JVI.75.22.10906-10911.2001
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Funding
- NIAID NIH HHS [R01 AI045463, UO1 AI49764, R01 AI39420, R01 AI039420, R01 AI45463] Funding Source: Medline
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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 and-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.
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