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TitleCryo-EM structures of antibodies elicited by germline-targeting HIV MPER epitope scaffolds.
Journal, issue, pagesCell Rep, Vol. 45, Issue 1, Page 116818, Year 2026
Publish dateJan 27, 2026
AuthorsJiachen Huang / Olivia M Swanson / Kimmo Rantalainen / Monica L Fernández-Quintero / Johannes R Loeffler / Ryan Tingle / Erik Georgeson / Nicole Phelps / Gabriel Ozorowski / Torben Schiffner / William R Schief / Andrew B Ward /
PubMed AbstractApplying cryoelectron microscopy (cryo-EM) to small protein complexes is usually challenging due to their lack of features for particle alignment. Here, we characterized antibody responses to 21 kDa ...Applying cryoelectron microscopy (cryo-EM) to small protein complexes is usually challenging due to their lack of features for particle alignment. Here, we characterized antibody responses to 21 kDa human immunodeficiency virus (HIV) membrane-proximal external region germline-targeting (MPER-GT) immunogens through cryo-EM by complexing them with 10E8 or Fabs derived from MPER-GT-immunized animals. Distinct antibody-antigen interactions were analyzed using atomic models generated from cryo-EM maps. Mutagenesis screening revealed that off-target monoclonal antibodies (mAbs), which do not compete with 10E8, bind non-MPER epitopes, and the binding of two most dominant epitopes were verified by cryo-EM. The structures of 10E8-class on-target Fabs showed binding patterns that resemble the YxFW motif in the 10E8 heavy chain complementarity-determining region 3 (HCDR3) loop. Additionally, we demonstrate that high-resolution maps can be generated from heterogeneous samples with pooled competing Fabs. Overall, our findings will facilitate the optimization of MPER-GT antigens and push the size limit for cryo-EM-based epitope mapping with smaller antigens and heterogeneous antibody mixes.
External linksCell Rep / PubMed:41528847
MethodsEM (single particle)
Resolution4.2 - 6.3 Å
Structure data

EMDB-71884: 10E8-GT10.2 immunogen in complex with mouse Fab W6-10 and rhesus macaque Fab W10-09
Method: EM (single particle) / Resolution: 6.3 Å

EMDB-71885: 10E8-GT10.2 immunogen in complex with Rhesus macaque Fab W3-02 and mouse Fab W6-03
Method: EM (single particle) / Resolution: 4.2 Å

Source
  • Mus musculus (house mouse)
  • synthetic construct (others)
  • Homo sapiens (human)
  • Macaca mulatta (Rhesus monkey)

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