C OMMUNICATION Inhibition of Mammalian Glycan Biosynthesis Produces Non-self Antigens for a Broadly Neutralising, HIV-1 Specific Antibody Christopher N. Scanlan 1 ⁎†, Gayle E. Ritchie 1 †, Kavitha Baruah 2 Max Crispin 1 , David J. Harvey 1 , Bernhard B. Singer 3 , Lothar Lucka 4 Mark R. Wormald 1 , Paul Wentworth Jr 2 , Nicole Zitzmann 1 Pauline M. Rudd 5 , Dennis R. Burton 6 and Raymond A. Dwek 1 1 Glycobiology Institute, Department of Biochemistry , University of Oxford, Oxford OX1 3QU 2 Scripps-Oxford Laboratory , Department of Biochemistry , University of Oxford, Oxford OX1 3QU 3 Institute of Anatomy , University Hospital Essen, Hufelandstr. 55, 45122 Essen, Germany 4 Institute of Biochemistry and Molecular Biology, Charite- Universitätsmedizin Berlin, Campus Benjamin Franklin, Arnimallee 22, D-14195 Berlin, Germany 5 National Institute for Bioprocessing Research and Training, Conway Institute, University College Dublin, Belfield, Dublin 4 6 Department of Immunology , Scripps Research Institute, 10550 North Torry Pines Road, La Jolla, CA 92037, USA The HIV envelope has evolved a dense array of immunologically “self” carbohydrates that efficiently protect the virus from antibody recognition. Nonetheless, one broadly neutralising antibody, IgG1 2G12, has been shown to recognise a cluster of oligomannose glycans on the HIV-1 surface antigen gp120. Thus the self carbohydrates of HIV are now regarded as potential targets for viral neutralisation and vaccine design. Here, we show that chemical inhibition of mammalian glycoprotein synthesis, with the plant alkaloid kifunensine, creates multiple HIV (2G12) epitopes on the surface of previously non-antigenic self proteins and cells, including HIV gp120. This formally demonstrates the structural basis for self/non-self discrimination between viral and host glycans, by a neutralising antibody. Moreover, this study provides an alternative protein engineering approach to the design of a carbohydrate vaccine for HIV-1 by chemical synthesis. © 2007 Elsevier Ltd. All rights reserved. *Corresponding author Keywords: human immunodeficiency virus; neutralising antibody; vaccine; 2G12 † These two authors contributed equally to this work. Abbreviations used: ESI, electrospray ionization; MALDI, matrix-assisted laser desorption/ionization; Man, mannose; MS, mass spectrometry; Q, quadrupole; RT-PCR, reverse transcriptase polymerase chain reaction; TOF, time-of-flight. E-mail address of the corresponding author: chris.scanlan@bioch.ox.ac.uk doi:10.1016/j.jmb.2007.06.027 J. Mol. Biol. (2007) 372, 16–22 0022-2836/$ - see front matter © 2007 Elsevier Ltd. All rights reserved.