Journal: eBioMedicine

Article Title: Somatic hypermutation shapes the viral escape profile of SARS-CoV-2 neutralising antibodies

doi: 10.1016/j.ebiom.2025.105770

Figure Lengend Snippet: Hamsters treated with SARS-CoV-2 neutralising mAbs and subsequently infected with SARS-CoV-2 occasionally show breakthrough infections. a) Schematic depiction of the experimental outline. b) Virus titres in lungs and nasal turbinates of SARS-CoV-2 infected animals at 4 dpi. c) Total lung scores for pathomorphological changes in hamsters at 4 dpi. d) Pathomorphological changes of hamster lungs. Representative images of one animal per treatment group (median group score) are shown at 4 dpi. The treatment group is indicated above. Inserts represent higher magnifications of the respective lungs. While no or minimal lesions were identified in all three mAbs treatment groups (MB025_A07K, MB025_A09L, MB027_E04L), multifocal atelectasis (asterisk) with prominent perivascular and peribronchial inflammation (arrows) as well as perivascular oedema (arrow heads) was present in the Palivizumab treated hamsters (V: vessel, B: bronchiole). Haematoxylin and eosin staining. e) SARS-CoV-2 genome sequencing data for the three breakthrough infections and an infected animal from the control group (Palivizumab). At each nucleotide position (X-axis), the plot shows the frequency of A, T, G, and C as detected by Oxford Nanopore sequencing. The cutoff of 10% was chosen to discriminate sequencing noise from specific signal. For non-synonymous mutations, the consequential amino acid exchange is indicated as dot label. The mutation G485D is highlighted in red.

Article Snippet: Fast in vitro viral antibody escape assay reveals spike mutations that prevent mAb binding. a) Experimental outline: Recombinant, replication-competent VSV∗ΔG-S was used to select escape mutants in the presence of mAbs. b) Exemplary result for the Sanger sequencing of untreated virus (upper panel) and virus treated with a control mAb REGN10987 (lower panel). c) Mutation frequency determination of control mAb REGN10987 using a customised Oxford Nanopore sequencing pipeline.

Techniques: Infection, Virus, Staining, Sequencing, Control, Nanopore Sequencing, Mutagenesis

Journal: eBioMedicine

Article Title: Somatic hypermutation shapes the viral escape profile of SARS-CoV-2 neutralising antibodies

doi: 10.1016/j.ebiom.2025.105770

Figure Lengend Snippet: Fast in vitro viral antibody escape assay reveals spike mutations that prevent mAb binding. a) Experimental outline: Recombinant, replication-competent VSV∗ΔG-S was used to select escape mutants in the presence of mAbs. b) Exemplary result for the Sanger sequencing of untreated virus (upper panel) and virus treated with a control mAb REGN10987 (lower panel). c) Mutation frequency determination of control mAb REGN10987 using a customised Oxford Nanopore sequencing pipeline. At each nucleotide position (X-axis), the plot shows frequencies of A, T, G, and C as detected by Oxford Nanopore sequencing. The cutoff of 10% was chosen to discriminate sequencing noise from specific signal. For non-synonymous mutations, the consequential amino acid exchange is indicated as dot label. d) Sequencing results of escape mutants that were generated against a panel of SARS-CoV-2 neutralising mAbs which were described previously. e) Somatic hypermutation analysis of mAb MB025_A07K and determination of viral escape mutants selected against the germline and somatically hypermutated mAb versions. V gene segments are indicated in regular fonts with SHM highlighted in red. D and J gene segments are indicated by italic fonts, no SHM were altered in these regions. Complementarity-determining regions (CDR) as determined by IgBlast are highlighted in grey boxes. f) Similar analysis as in e) for the mAb MB025_A09L.

Article Snippet: Fast in vitro viral antibody escape assay reveals spike mutations that prevent mAb binding. a) Experimental outline: Recombinant, replication-competent VSV∗ΔG-S was used to select escape mutants in the presence of mAbs. b) Exemplary result for the Sanger sequencing of untreated virus (upper panel) and virus treated with a control mAb REGN10987 (lower panel). c) Mutation frequency determination of control mAb REGN10987 using a customised Oxford Nanopore sequencing pipeline.

Techniques: In Vitro, Binding Assay, Recombinant, Sequencing, Virus, Control, Mutagenesis, Nanopore Sequencing, Generated