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TitleStructural and functional evaluation of de novo-designed, two-component nanoparticle carriers for HIV Env trimer immunogens.
Journal, issue, pagesPLoS Pathog, Vol. 16, Issue 8, Page e1008665, Year 2020
Publish dateAug 11, 2020
AuthorsAleksandar Antanasijevic / George Ueda / Philip J M Brouwer / Jeffrey Copps / Deli Huang / Joel D Allen / Christopher A Cottrell / Anila Yasmeen / Leigh M Sewall / Ilja Bontjer / Thomas J Ketas / Hannah L Turner / Zachary T Berndsen / David C Montefiori / Per Johan Klasse / Max Crispin / David Nemazee / John P Moore / Rogier W Sanders / Neil P King / David Baker / Andrew B Ward /
PubMed AbstractTwo-component, self-assembling nanoparticles represent a versatile platform for multivalent presentation of viral antigens. Computational design of protein nanoparticles with differing sizes and ...Two-component, self-assembling nanoparticles represent a versatile platform for multivalent presentation of viral antigens. Computational design of protein nanoparticles with differing sizes and geometries enables combination with antigens of choice to test novel multimerization concepts in immunization strategies where the goal is to improve the induction and maturation of neutralizing antibody lineages. Here, we describe detailed antigenic, structural, and functional characterization of computationally designed tetrahedral, octahedral, and icosahedral nanoparticle immunogens displaying trimeric HIV envelope glycoprotein (Env) ectodomains. Env trimers, based on subtype A (BG505) or consensus group M (ConM) sequences and engineered with SOSIP stabilizing mutations, were fused to an underlying trimeric building block of each nanoparticle. Initial screening yielded one icosahedral and two tetrahedral nanoparticle candidates, capable of presenting twenty or four copies of the Env trimer. A number of analyses, including detailed structural characterization by cryo-EM, demonstrated that the nanoparticle immunogens possessed the intended structural and antigenic properties. When the immunogenicity of ConM-SOSIP trimers presented on a two-component tetrahedral nanoparticle or as soluble proteins were compared in rabbits, the two immunogens elicited similar serum antibody binding titers against the trimer component. Neutralizing antibody titers were slightly elevated in the animals given the nanoparticle immunogen and were initially more focused to the trimer apex. Altogether, our findings indicate that tetrahedral nanoparticles can be successfully applied for presentation of HIV Env trimer immunogens; however, the optimal implementation to different immunization strategies remains to be determined.
External linksPLoS Pathog / PubMed:32780770 / PubMed Central
MethodsEM (single particle)
Resolution4.14 - 19.68 Å
Structure data

EMDB-21175:
EM-Based Polyclonal Epitope Mapping. ConM-SOSIP Complexed with Purified Polyclonal Fab (Grp 1, ID r2381 Wk4)
Method: EM (single particle) / Resolution: 17.24 Å

EMDB-21176:
EM-Based Polyclonal Epitope Mapping. ConM-SOSIP Complexed with Purified Polyclonal Fab (Grp 1, ID r2382, Wk4)
Method: EM (single particle) / Resolution: 16.52 Å

EMDB-21177:
EM-Based Polyclonal Epitope Mapping. ConM-SOSIP Complexed with Purified Polyclonal Fab (Grp 2, ID r2383 Wk4)
Method: EM (single particle) / Resolution: 17.23 Å

EMDB-21178:
EM-Based Polyclonal Epitope Mapping. ConM-SOSIP Complexed with Purified Polyclonal Fab (Grp 2, ID r2385 Wk4)
Method: EM (single particle) / Resolution: 18.02 Å

EMDB-21179:
EM-Based Polyclonal Epitope Mapping. ConM-SOSIP Complexed with Purified Polyclonal Fab (Grp 1, ID r2381 Wk22)
Method: EM (single particle) / Resolution: 17.89 Å

EMDB-21180:
EM-Based Polyclonal Epitope Mapping. ConM-SOSIP Complexed with Purified Polyclonal Fab (Grp 1, ID r2382 Wk22)
Method: EM (single particle) / Resolution: 19.68 Å

EMDB-21181:
EM-Based Polyclonal Epitope Mapping. ConM-SOSIP Complexed with Purified Polyclonal Fab (Grp 2, ID r2383 Wk22)
Method: EM (single particle) / Resolution: 17.11 Å

EMDB-21182:
EM-Based Polyclonal Epitope Mapping. ConM-SOSIP Complexed with Purified Polyclonal Fab (Grp 2, ID r2385 Wk22)
Method: EM (single particle) / Resolution: 17.11 Å

EMDB-21183:
De novo designed icosahedral nanoparticle I53_dn5 presenting BG505-SOSIP, Cryo-EM Map
Method: EM (single particle) / Resolution: 12.46 Å

EMDB-21184:
BG505-SOSIP map reconstructed by subparticle extraction and refinement from an icosahedral nanoparticle (I53_dn5)
Method: EM (single particle) / Resolution: 6.68 Å

21185

6vfk

EMDB-21185, PDB-6vfk:
De novo designed tetrahedral nanoparticle T33_dn10 displaying 4 copies of BG505-SOSIP trimer on the surface
Method: EM (single particle) / Resolution: 4.25 Å

21186

6vfl

EMDB-21186, PDB-6vfl:
BG505-SOSIP model reconstructed by subparticle extraction and refinement from a tetrahedral nanoparticle T33_dn10
Method: EM (single particle) / Resolution: 4.14 Å

Chemicals

ChemComp-NAG:
2-acetamido-2-deoxy-beta-D-glucopyranose / N-Acetylglucosamine

Source
  • synthetic construct (others)
KeywordsDE NOVO PROTEIN / Nanoparticle / Rosetta / De novo protein design / HIV Env

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