The Chimpanzee SIV Envelope Trimer: Structure and Deployment as an HIV Vaccine Template

Andrabi, Raiees; Pallesen, Jesper; Allen, Joel D.; Song, Ge; Zhang, Jinsong; de Val, Natalia; Gegg, Gavin; Porter, Katelyn; Su, Ching-Yao; Pauthner, Matthias; Newman, Amanda; Bouton-Verville, Hilary; Garces, Fernando; Wilson, Ian A.; Crispin, Max; Hahn, Beatrice H.; Haynes, Barton F.; Verkoczy, Laurent; Ward, Andrew B.; Burton, Dennis R.

Detailed methods are provided in the online version of this paper and include the following: (2002) . PHENIX: building new software for automated crystallographic structure determination. Acta Crystallogr. D Biol. Crystallogr. 58, 1948 Crystallogr. 58, -1954 Agirre, J., Iglesias-Ferná ndez, J., Rovira, C., Davies, G.J., Wilson, K.S., and Cowtan, K.D. (2015) PEI-MAX 4000 transfection reagent (Polysciences, Inc.) . The secreted soluble trimers proteins were purified from cell supernatants after 5 days using agarose-bound Gallanthus Nivalis Lectin (GNL) (Vector Labs) or CNBr-activated Sepharose 4B bead (GE Healthcare) bound PGT145 bnAb antibody affinity columns as described previously (Pugach et al., 2015) . The affinity-purified proteins were size exclusion chromatography (SEC)-purified with a Superdex 200 10/300 GL column (GE Healthcare) in PBS/TBS. The purified trimers for the immunization experiments were quality control tested for antigenicity with a range of HIV Env-specific neutralizing and non-neutralizing mAbs.