Expression, Purification, and Isotope Labeling of a gp120 V3 Peptide and Production of a Fab from a HIV-1 Neutralizing Antibody for NMR Studies

2002 ◽  
Vol 24 (3) ◽  
pp. 374-383 ◽  
Author(s):  
Michal Sharon ◽  
Matthias Görlach ◽  
Rina Levy ◽  
Yehezkiel Hayek ◽  
Jacob Anglister
Keyword(s):  
Isotope Labeling ◽  
Neutralizing Antibody ◽  
Nmr Studies ◽  
Hiv 1

Expression, purification, and isotope labeling of the Fv of the human HIV-1 neutralizing antibody 447-52D for NMR studies

2003 ◽  
Vol 29 (2) ◽  
pp. 291-303 ◽  
Author(s):  
Naama Kessler ◽  
Anat Zvi ◽  
Min Ji ◽  
Michal Sharon ◽  
Osnat Rosen ◽  
...  
Keyword(s):  
Isotope Labeling ◽  
Neutralizing Antibody ◽  
Nmr Studies ◽  
Hiv 1

scholarly journals Large Multidomain Protein NMR: HIV-1 Reverse Transcriptase Precursor in Solution

2020 ◽  
Vol 21 (24) ◽  
pp. 9545
Author(s):  
Tatiana V. Ilina ◽  
Zhaoyong Xi ◽  
Teresa Brosenitsch ◽  
Nicolas Sluis-Cremer ◽  
Rieko Ishima
Keyword(s):  
Reverse Transcriptase ◽  
Isotope Labeling ◽  
Protein Nmr ◽  
Methyl Groups ◽  
Large Protein ◽  
Nmr Studies ◽  
Large Proteins ◽  
Domain Motion ◽  
Correlation Spectrum ◽  
Hiv 1

NMR studies of large proteins, over 100 kDa, in solution are technically challenging and, therefore, of considerable interest in the biophysics field. The challenge arises because the molecular tumbling of a protein in solution considerably slows as molecular mass increases, reducing the ability to detect resonances. In fact, the typical 1H-13C or 1H-15N correlation spectrum of a large protein, using a 13C- or 15N-uniformly labeled protein, shows severe line-broadening and signal overlap. Selective isotope labeling of methyl groups is a useful strategy to reduce these issues, however, the reduction in the number of signals that goes hand-in-hand with such a strategy is, in turn, disadvantageous for characterizing the overall features of the protein. When domain motion exists in large proteins, the domain motion differently affects backbone amide signals and methyl groups. Thus, the use of multiple NMR probes, such as 1H, 19F, 13C, and 15N, is ideal to gain overall structural or dynamical information for large proteins. We discuss the utility of observing different NMR nuclei when characterizing a large protein, namely, the 66 kDa multi-domain HIV-1 reverse transcriptase that forms a homodimer in solution. Importantly, we present a biophysical approach, complemented by biochemical assays, to understand not only the homodimer, p66/p66, but also the conformational changes that contribute to its maturation to a heterodimer, p66/p51, upon HIV-1 protease cleavage.

scholarly journals Myomedin scaffold variants targeted to 10E8 HIV-1 broadly neutralizing antibody mimic gp41 epitope and elicit HIV-1 virus-neutralizing sera in mice

Virulence ◽  
2021 ◽  
Vol 12 (1) ◽  
pp. 1271-1287
Author(s):  
Milan Kuchař ◽  
Petr Kosztyu ◽  
Veronika Daniel Lišková ◽  
Jiří Černý ◽  
Hana Petroková ◽  
...  
Keyword(s):  
Neutralizing Antibody ◽  
Broadly Neutralizing Antibody ◽  
Hiv 1

scholarly journals Immunogenicity of HIV-1-Based Virus-Like Particles with Increased Incorporation and Stability of Membrane-Bound Env

Vaccines ◽  
2021 ◽  
Vol 9 (3) ◽  
pp. 239
Author(s):  
Christopher A. Gonelli ◽  
Hannah A. D. King ◽  
Charlene Mackenzie ◽  
Secondo Sonza ◽  
Rob J. Center ◽  
...  
Keyword(s):  
Neutralizing Antibody ◽  
Antibody Responses ◽  
Neutralization Activity ◽  
Virus Like Particles ◽  
Antibody Isotypes ◽  
Immunodeficiency Virus ◽  
Proline Substitution ◽  
Membrane Bound ◽  
Antibody Epitopes ◽  
Hiv 1

An optimal prophylactic vaccine to prevent human immunodeficiency virus (HIV-1) transmission should elicit protective antibody responses against the HIV-1 envelope glycoprotein (Env). Replication-incompetent HIV-1 virus-like particles (VLPs) offer the opportunity to present virion-associated Env with a native-like structure during vaccination that closely resembles that encountered on infectious virus. Here, we optimized the incorporation of Env into previously designed mature-form VLPs (mVLPs) and assessed their immunogenicity in mice. The incorporation of Env into mVLPs was increased by replacing the Env transmembrane and cytoplasmic tail domains with those of influenza haemagglutinin (HA-TMCT). Furthermore, Env was stabilized on the VLP surface by introducing an interchain disulfide and proline substitution (SOSIP) mutations typically employed to stabilize soluble Env trimers. The resulting mVLPs efficiently presented neutralizing antibody epitopes while minimizing exposure of non-neutralizing antibody sites. Vaccination of mice with mVLPs elicited a broader range of Env-specific antibody isotypes than Env presented on immature VLPs or extracellular vesicles. The mVLPs bearing HA-TMCT-modified Env consistently induced anti-Env antibody responses that mediated modest neutralization activity. These mVLPs are potentially useful immunogens for eliciting neutralizing antibody responses that target native Env epitopes on infectious HIV-1 virions.

Regulation of the susceptibility of HIV-1 to a neutralizing antibody KD-247 by nonepitope mutations distant from its epitope

AIDS ◽  
2011 ◽  
Vol 25 (18) ◽  
pp. 2209-2216 ◽  
Author(s):  
Mari Takizawa ◽  
Kosuke Miyauchi ◽  
Emiko Urano ◽  
Shigeru Kusagawa ◽  
Katsuhiko Kitamura ◽  
...  
Keyword(s):  
Neutralizing Antibody ◽  
Hiv 1
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