|Entry||Database: EMDB / ID: EMD-9920|
|Title||The cryo-em structure of HPV18 VLP|
|Biological species||Human papillomavirus|
|Method||single particle reconstruction / cryo EM / Resolution: 11.5 Å|
|Authors||Li S / Zheng Q|
|Citation||Journal: Eur J Pharm Biopharm / Year: 2019|
Title: Altered antigenicity and immunogenicity of human papillomavirus virus-like particles in the presence of thimerosal.
Authors: Siyi Chen / Xiaofen Huang / Yike Li / Xin Wang / Huirong Pan / Zhijie Lin / Qingbing Zheng / Shaowei Li / Jun Zhang / Ningshao Xia / Qinjian Zhao /
Abstract: Thimerosal has been widely used as a preservative in human vaccines for decades. Thimerosal, a thiol capping agent with ethyl mercury being the active degradant, could have impacts on the vaccine ...Thimerosal has been widely used as a preservative in human vaccines for decades. Thimerosal, a thiol capping agent with ethyl mercury being the active degradant, could have impacts on the vaccine potency due to potential thiol modification. The effects on the antigenicity and immunogenicity of human papillomavirus (HPV) virus-like particles (VLPs) in the presence of thimerosal was studied. In general, reduced binding activity was observed between HPV antigens and monoclonal antibodies (mAbs) upon thimerosal treatment, accompanied by reduced protein conformational stability. The immunogenicity of a pentavalent vaccine formulation (HPV6, HPV11, HPV16, HPV18 and hepatitis E virus) with or without thimerosal was studied in mice. The functional antibody titres, as well as the binding titres, were determined, showing a substantial decrease for vaccine formulations containing thimerosal for HPV16/18. Similarly, epitope-specific competition assays using specific and functional mAbs as tracers also showed a significant reduction in immunogenicity for HPV16/18 in the presence of thimerosal. Structural alterations in the capsid protein for HPV18 were observed with cryo-electron microscopy and 3-dimensional reconstruction in the comparative structural analysis. The results should alert scientists in formulation development field on the choice for vaccine preservatives, in particular for thiol-containing antigens.
|Structure viewer||EM map: |
Downloads & links
|File||Download / File: emd_9920.map.gz / Format: CCP4 / Size: 1.9 GB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)|
|Projections & slices|
Images are generated by Spider.
|Voxel size||X=Y=Z: 1.128 Å|
|Symmetry||Space group: 1|
CCP4 map header:
-Entire Human papillomavirus
|Entire||Name: Human papillomavirus / Number of components: 1|
-Component #1: virus, Human papillomavirus
|Virus||Name: Human papillomavirusHuman papillomavirus infection / Class: VIRUS-LIKE PARTICLE / Empty: Yes / Enveloped: No / Isolate: OTHER|
|Species||Species: Human papillomavirus|
|Source (engineered)||Expression System: Escherichia coli-Pichia pastoris shuttle vector pPpARG4 (others)|
|Specimen||Specimen state: Particle / Method: cryo EM|
|Sample solution||Specimen conc.: 2 mg/mL / pH: 7.4|
|Vitrification||Instrument: FEI VITROBOT MARK IV / Cryogen name: ETHANE / Humidity: 100 %|
-Electron microscopy imaging
Model: Tecnai F30 / Image courtesy: FEI Company
|Imaging||Microscope: FEI TECNAI F30|
|Electron gun||Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Electron dose: 25 e/Å2 / Illumination mode: FLOOD BEAM|
|Lens||Imaging mode: BRIGHT FIELD|
|Specimen Holder||Model: GATAN LIQUID NITROGEN|
|Camera||Detector: FEI FALCON III (4k x 4k)|
|Processing||Method: single particle reconstruction / Applied symmetry: I (icosahedral) / Number of projections: 901|
|3D reconstruction||Resolution: 11.5 Å / Resolution method: FSC 0.143 CUT-OFF|
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