1. Volume 21, Issue 10, Pages 2992-3002 (December 2017)
Soluble Prefusion Closed DS-SOSIP.664-Env Trimers of Diverse HIV-1 Strains 
M. Gordon Joyce, Ivelin S. Georgiev, Yongping Yang, Aliaksandr Druz, Hui Geng, Gwo-Yu Chuang, Young Do Kwon, Marie Pancera, Reda Rawi, Mallika Sastry, Guillaume B.E. Stewart-Jones, Angela Zheng, Tongqing Zhou, Misook Choe, Joseph G. Van Galen, Rita E. Chen, Christopher R. Lees, Sandeep Narpala, Michael Chambers, Yaroslav Tsybovsky, Ulrich Baxa, Adrian B. McDermott, John R. Mascola, Peter D. Kwong 
Cell Reports 
Volume 21, Issue 10, Pages (December 2017)
DOI: /j.celrep
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2. Cell Reports 2017 21, 2992-3002DOI: (10.1016/j.celrep.2017.11.016)
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3. Figure 1 Design of Chimeric gp120 BG505 DS-SOSIP HIV-1-Env Trimers
(A) Ribbon diagram of BG505.SOSIP-Env trimer in its closed prefusion state with gp120 subunit in light blue and gp41 subunit in light red, with inset of the gp41 wrapping around the N and C termini of gp120.
(B) Surface representation of HIV-1-Env in its closed prefusion state with gp120 in light blue and gp41 in salmon.
(C) Schematic of gp120 and gp41 sequences with SOS, I559P, and DS mutations, along with a second schematic of a gp120 chimera with non-BG505 region colored in purple.
(D) Surface representation of gp120 chimera SOSIP.DS molecules with purple for other gp120 strains, salmon for BG505.SOSIP.664 gp41, and light blue for BG505 gp120 region.
(E) Surface representation of gp120 chimera for SC422.8 clade B Env (left) and ZM106.9 clade C Env (right).
See also Figure S1 and Table S1.
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4. Figure 2
Antigenic Selection of Prefusion Closed HIV-1-Env Trimers with DS-SOSIP or BG505 DS-SOSIP Chimera Design
(A) Schematic of the high-throughput 96-well expression-antigenic screening system.
(B) Antibodies used in antigenic selection, rationale for use, and antibody neutralization breadth on the 180-isolate panel.
(C) HIV-Env trimer-binding responses relative to BG505 DS-SOSIP. The values displayed at the top of the graph indicates the number of Env molecules with binding response at least 75% that of BG505 DS-SOSIP Env. Dotted line indicates 75% binding response relative to BG505 DS-SOSIP.
See also Tables S2–S4 and Figures S4 and S5.
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5. Figure 3 Properties of DS-SOSIP.664 from Clade C Strain ZM106.9
(A) Gel filtration profile for chimeric ZM106.9 DS-SOSIP.664 obtained following GNA-lectin affinity chromatography.
(B) SDS-PAGE analysis of purified chimeric ZM106.9 DS-SOSIP. A diffuse band corresponding to glycosylated gp41 can be seen under reducing conditions. M, markers; 1, ZM106.9 chimeric DS-SOSIP.664 under non-reducing conditions; 2, ZM106.9 chimeric DS-SOSIP.664 under reducing conditions.
(C) Antigenic analysis of ZM106.9 DS-SOSIP.664 following D negative selection (data shown are the mean of two independent assays).
See also Table S3.
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6. Figure 4
Properties of DS-SOSIP.664 Chimera from Clade C Strain 3301.V1.C24
(A) Gel filtration profile for chimeric 3301.V1.C24 DS-SOSIP.664 chimera obtained from lectin affinity chromatography.
(B) SDS-PAGE analysis of purified 3301.V1.C24 DS-SOSIP chimera. A somewhat diffuse band corresponding to glycosylated gp41 can be seen under reducing conditions. M, markers; 1, 3301.V1.C24 DS-SOSIP.664 chimera under non-reducing conditions; 2, 3301.V1.C24 DS-SOSIP.664 chimera under reducing conditions.
(C) Antigenic analysis of 3301.V1.C24 DS-SOSIP.664 chimera following D negative selection (data shown are the mean of two independent assays).
See also Table S3.
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7. Figure 5 Properties of DS-SOSIP.664 from Clade C Strain 25925.22
(A) Gel filtration profile for chimeric DS-SOSIP.664 chimera obtained from lectin affinity chromatography.
(B) SDS-PAGE analysis of purified chimeric DS-SOSIP. A diffuse band corresponding to glycosylated gp41 can be seen under reducing conditions. M, markers; 1, DS-SOSIP.664 chimera under non-reducing conditions; 2, DS-SOSIP.664 chimera under reducing conditions.
(C) Antigenic analysis of DS-SOSIP.664 chimera following D negative selection (data shown are the mean of two independent assays).
See also Table S3.
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8. Figure 6
Negative-Stain EM of Selected DS-SOSIP.664 Chimera Env Trimer Molecules
EM analysis was performed on proteins after lectin affinity and size exclusion chromatography. Shown are images of class averages for five of the chimeric gp140 constructs from different HIV-1 strains. The scale bar represents 5 nm.
See also Figure S3.
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9. Figure 7
HIV-1 Strains that Form Closed Prefusion Molecules in Either the SOSIP or DS-SOSIP Chimera Designs
(A) HIV-1-Env molecules from different strains that could form closed prefusion Env trimer molecules are displayed on a representative HIV-1 phylogenetic tree constructed using global isolate sequences of HIV-1.
(B) A set of 19 HIV-1 strains from different clades was also assessed for alternate design strategies, including incorporation of additional interface residues (46–54, 70–75, 84–89, 99, 102, 106, 107, 114, 215, 220–224, 226, 244, 471–473, and 476–477) from BG505.SOSIP (DS-SOSIP chimera-Int), as well as use of a 15-amino acid linker region combined with the DS-SOSIP chimera design (DS-SOSIP chimera.sc15ln). Dotted line indicates 75% binding response relative to BG505 DS-SOSIP.
See also Tables S1, S3, and S4.
Cell Reports  , DOI: ( /j.celrep )
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