Engineering and characterization of a fluorescent native-like HIV-1 envelope glycoprotein trimer

Generation of a stable, soluble mimic of the HIV-1 envelope glycoprotein (Env) trimer on the virion surface has been considered an important first step for developing a successful HIV-1 vaccine. Recently, a soluble native-like Env trimer (BG505 SOSIP.664) has been described. This protein has facilit...

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Main Authors: Pritchard, L, Crispin, M, Sliepen, K, van Montfort, T, Ozorowski, G, Ward, A, Sanders, R
Format: Journal article
Published: MDPI 2015
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author Pritchard, L
Crispin, M
Sliepen, K
van Montfort, T
Ozorowski, G
Ward, A
Sanders, R
author_facet Pritchard, L
Crispin, M
Sliepen, K
van Montfort, T
Ozorowski, G
Ward, A
Sanders, R
author_sort Pritchard, L
collection OXFORD
description Generation of a stable, soluble mimic of the HIV-1 envelope glycoprotein (Env) trimer on the virion surface has been considered an important first step for developing a successful HIV-1 vaccine. Recently, a soluble native-like Env trimer (BG505 SOSIP.664) has been described. This protein has facilitated major advances in the HIV-1 vaccine field, since it was the first Env immunogen that induced consistent neutralizing antibodies against a neutralization-resistant (tier 2) virus. Moreover, BG505 SOSIP.664 enabled elucidation of the atomic resolution structure of the Env trimer and facilitated the isolation and characterization of new broadly neutralizing antibodies against HIV-1. Here, we designed and characterized the BG505 SOSIP.664 trimer fused to fluorescent superfolder GFP (sfGFP), a GFP variant that allows efficient folding (BG505 SOSIP.664-sfGFP). Despite the presence of the sfGFP, the Env protein largely retained its morphology, antigenicity, glycan composition, and thermostability. In addition, we show that BG505 SOSIP.664-sfGFP can be used for fluorescence-based assays, such as flow cytometry.
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spelling oxford-uuid:f32f0eaf-6ec4-4e32-b17f-14c8763d6d692022-03-27T12:10:08ZEngineering and characterization of a fluorescent native-like HIV-1 envelope glycoprotein trimerJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:f32f0eaf-6ec4-4e32-b17f-14c8763d6d69Symplectic Elements at OxfordMDPI2015Pritchard, LCrispin, MSliepen, Kvan Montfort, TOzorowski, GWard, ASanders, RGeneration of a stable, soluble mimic of the HIV-1 envelope glycoprotein (Env) trimer on the virion surface has been considered an important first step for developing a successful HIV-1 vaccine. Recently, a soluble native-like Env trimer (BG505 SOSIP.664) has been described. This protein has facilitated major advances in the HIV-1 vaccine field, since it was the first Env immunogen that induced consistent neutralizing antibodies against a neutralization-resistant (tier 2) virus. Moreover, BG505 SOSIP.664 enabled elucidation of the atomic resolution structure of the Env trimer and facilitated the isolation and characterization of new broadly neutralizing antibodies against HIV-1. Here, we designed and characterized the BG505 SOSIP.664 trimer fused to fluorescent superfolder GFP (sfGFP), a GFP variant that allows efficient folding (BG505 SOSIP.664-sfGFP). Despite the presence of the sfGFP, the Env protein largely retained its morphology, antigenicity, glycan composition, and thermostability. In addition, we show that BG505 SOSIP.664-sfGFP can be used for fluorescence-based assays, such as flow cytometry.
spellingShingle Pritchard, L
Crispin, M
Sliepen, K
van Montfort, T
Ozorowski, G
Ward, A
Sanders, R
Engineering and characterization of a fluorescent native-like HIV-1 envelope glycoprotein trimer
title Engineering and characterization of a fluorescent native-like HIV-1 envelope glycoprotein trimer
title_full Engineering and characterization of a fluorescent native-like HIV-1 envelope glycoprotein trimer
title_fullStr Engineering and characterization of a fluorescent native-like HIV-1 envelope glycoprotein trimer
title_full_unstemmed Engineering and characterization of a fluorescent native-like HIV-1 envelope glycoprotein trimer
title_short Engineering and characterization of a fluorescent native-like HIV-1 envelope glycoprotein trimer
title_sort engineering and characterization of a fluorescent native like hiv 1 envelope glycoprotein trimer
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