Matteo Bianchi / Hannah L Turner / Bartek Nogal / Christopher A Cottrell / David Oyen / Matthias Pauthner / Raiza Bastidas / Rebecca Nedellec / Laura E McCoy / Ian A Wilson / Dennis R Burton / Andrew B Ward / Lars Hangartner /
PubMed Abstract
Characterizing polyclonal antibody responses via currently available methods is inherently complex and difficult. Mapping epitopes in an immune response is typically incomplete, which creates a ...Characterizing polyclonal antibody responses via currently available methods is inherently complex and difficult. Mapping epitopes in an immune response is typically incomplete, which creates a barrier to fully understanding the humoral response to antigens and hinders rational vaccine design efforts. Here, we describe a method of characterizing polyclonal responses by using electron microscopy, and we applied this method to the immunization of rabbits with an HIV-1 envelope glycoprotein vaccine candidate, BG505 SOSIP.664. We detected known epitopes within the polyclonal sera and revealed how antibody responses evolved during the prime-boosting strategy to ultimately result in a neutralizing antibody response. We uncovered previously unidentified epitopes, including an epitope proximal to one recognized by human broadly neutralizing antibodies as well as potentially distracting non-neutralizing epitopes. Our method provides an efficient and semiquantitative map of epitopes that are targeted in a polyclonal antibody response and should be of widespread utility in vaccine and infection studies.
EMDB-7552: CryoEM map of BG505 SOSIP.664 in complex with post boost 1 serum from rabbit 3417 Method: EM (single particle) / Resolution: 4.7 Å
EMDB-7553: CryoEM map of BG505 SOSIP.664 in complex with post boost 1 serum from rabbit 3417 Method: EM (single particle) / Resolution: 4.7 Å
EMDB-7554: CryoEM map of BG505 SOSIP.664 in complex with post boost 1 serum from rabbit 3417 Method: EM (single particle) / Resolution: 4.7 Å
EMDB-7555: CryoEM map of BG505 SOSIP.664 in complex with post boost 1 serum from rabbit 3417 Method: EM (single particle) / Resolution: 4.7 Å
EMDB-7556: CryoEM map of BG505 SOSIP.664 in complex with post boost 1 serum from rabbit 3417 Method: EM (single particle) / Resolution: 4.7 Å
EMDB-7557: CryoEM map of BG505 SOSIP.664 in complex with post boost 1 serum from rabbit 3417 Method: EM (single particle) / Resolution: 4.7 Å
EMDB-7570: Negative stain EM map of BG505 SOSIP.644 in complex with PG9 and 12N Fabs Method: EM (single particle) / Resolution: 17.36 Å
EMDB-7887: Negative Stain EM map of polyclonal serum in complex with BG505 SOSIP.664 from rabbit 3418 at post boost 2 Method: EM (single particle) / Resolution: 20.0 Å
EMDB-7888: Negative Stain EM map of polyclonal serum in complex with BG505 SOSIP.664 from rabbit 3418 at post boost 2. Method: EM (single particle) / Resolution: 20.0 Å
EMDB-7889: Negative Stain EM map of polyclonal serum in complex with BG505 SOSIP.664 from rabbit 3419 at post boost 2. Method: EM (single particle) / Resolution: 20.0 Å
EMDB-7890: Negative Stain EM map of polyclonal serum in complex with BG505 SOSIP.664 from rabbit 3419 at post boost 2. Method: EM (single particle) / Resolution: 20.0 Å
EMDB-7891: Negative Stain EM map of polyclonal serum in complex with BG505 SOSIP.664 from rabbit 3417 at post boost 2. Method: EM (single particle) / Resolution: 20.0 Å
EMDB-7892: Negative Stain EM map of polyclonal serum in complex with BG505 SOSIP.664 from rabbit 3417 at post boost 2. Method: EM (single particle) / Resolution: 20.0 Å
EMDB-7893: Negative Stain EM map of polyclonal serum in complex with BG505 SOSIP.664 from rabbit 3417 at post boost 2. Method: EM (single particle) / Resolution: 20.0 Å
EMDB-7894: Negative Stain EM map of polyclonal serum in complex with BG505 SOSIP.664 from rabbit 3420 at post boost 2. Method: EM (single particle) / Resolution: 20.0 Å
EMDB-7895: Negative Stain EM map of polyclonal serum in complex with BG505 MD39 CPG9 from rabbit 3417 at post boost 3 Method: EM (single particle) / Resolution: 20.0 Å
EMDB-7896, PDB-6did: HIV Env BG505 SOSIP with polyclonal Fabs from immunized rabbit #3417 post-boost#1 Method: EM (single particle) / Resolution: 4.71 Å
EMDB-7903: Negative stain EM map of BG505 SOSIP.644 in complex with PGT121 and 12N Fabs Method: EM (single particle) / Resolution: 17.36 Å
EMDB-7904: Negative stain EM map of BG505 SOSIP.644 in complex with PGT121 and serum from rabbit 3417 PB1 Method: EM (single particle) / Resolution: 18.45 Å
EMDB-7906: Negative stain EM map of BG505 SOSIP.644 in complex with PG9 Fab and serum from rabbit 3417 PB1 Method: EM (single particle) / Resolution: 18.45 Å
PDB-6cjk: Anti HIV Fab 10A Method: X-RAY DIFFRACTION / Resolution: 1.795 Å
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