2019-05-15RELRCSBRCSBRCSB2018-03-132018-04-112019-01-232019-05-15CryoEM map of BG505 SOSIP.664 in complex with post boost 1 serum from rabbit 3417Turner HLWard ABNogal BBianchi MTurner HLNogal BCottrell CAOyen DPauthner MBastidas RNedellec RMcCoy LEWilson IABurton DRWard ABHangartner LElectron-Microscopy-Based Epitope Mapping Defines Specificities of Polyclonal Antibodies Elicited during HIV-1 BG505 Envelope Trimer Immunization.ImmunityUS49288300.e8201830097292doi:10.1016/j.immuni.2018.07.0091097-41802048IUNIEHEMD-7557associated EM volumeComplex of BG505 SOSIP.664 and serum antibodies from post boost 1 from rabbit 3417Complex of BG505 SOSIP.664 and serum antibodies from post boost 1 from rabbit 34170Serum was purified through SEC after using a Protein A or G column to capture complex then cleave IgG releasing Fab and trimer. Substoichiometric occupancy and flexibility show lack of density in Fab region while trimer core is refined to higher resolution. Map shows two strong densities for Fabs bound to the glycan hole and partial densities near the base of the trimer.Human immunodeficiency virusHomo sapiensHEK293FsingleParticleparticle1.447.4TBSQuantifoil R1.2/1.3ETHANE100FEI VITROBOT MARK IVSample in TBS, added 0.5uL of DDM at 0.42mM. Sample was monodisperseFEI TECNAI ARCTICAFLOOD BEAMBRIGHT FIELDFIELD EMISSION GUN200FEI TITAN KRIOS AUTOGRID HOLDERNITROGENGATAN K2 SUMMIT (4k x 4k)COUNTING1-4611.560.01DogPicker was used to pick particles from micrographsCTFFIND35I8H1C1FOURIER SPACE4.7FSC 0.143 CUT-OFFcryoSPARC0.6.310828ANGULAR RECONSTITUTIONRELION2.1ANGULAR RECONSTITUTIONcryoSPARC0.6.3