Sliepen Kwinten, van Montfort Thijs, Ozorowski Gabriel, Pritchard Laura K, Crispin Max, Ward Andrew B, Sanders Rogier W
Department of Medical Microbiology, Academic Medical Center, University of Amsterdam, Amsterdam 1105 AZ, The Netherlands.
Department of Integrative Structural and Computational Biology, IAVI Neutralizing Antibody Center, Collaboration for AIDS Vaccine Discovery (CAVD), Center for HIV/AIDS Vaccine Immunology and Immunogen Discovery, The Scripps Research Institute, La Jolla, CA 92037, USA.
Biomolecules. 2015 Oct 23;5(4):2919-34. doi: 10.3390/biom5042919.
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.
在病毒粒子表面生成稳定、可溶的HIV-1包膜糖蛋白（Env）三聚体模拟物被认为是开发成功的HIV-1疫苗的重要第一步。最近，一种可溶性天然样Env三聚体（BG505 SOSIP.664）已被报道。这种蛋白推动了HIV-1疫苗领域的重大进展，因为它是第一种能诱导针对中和抗性（2级）病毒产生一致中和抗体的Env免疫原。此外，BG505 SOSIP.664使得能够阐明Env三聚体的原子分辨率结构，并有助于分离和鉴定针对HIV-1的新型广谱中和抗体。在此，我们设计并表征了与荧光超折叠绿色荧光蛋白（sfGFP，一种允许高效折叠的GFP变体）融合的BG505 SOSIP.664三聚体（BG505 SOSIP.664-sfGFP）。尽管存在sfGFP，Env蛋白在很大程度上保留了其形态、抗原性、聚糖组成和热稳定性。此外，我们表明BG505 SOSIP.664-sfGFP可用于基于荧光的检测，如流式细胞术。
