| Products/Services Used | Details | Operation |
|---|---|---|
| Gene Synthesis | Mutations found in the most enriched clones were incorporated into the most recent designs, synthesized at Genscript either as C-terminal His tagged gp120s or MD39/MD64 trimers in the pHLsec vector and expressed and purified as described previously (22)...To obtain multivalent immunogens, trimers were genetically fused to ferritin from Helicobacter pylori using a short flexible linker. Genes were codon optimized for HEK293 cells and cloned into the pHLsec plasmid (GenScript)...mAb binding ELISAs were performed by capturing antigen (1 µg/ml) onto plates precoated with anti-His antibody (1 µg/ml; Genscript) and blocked with blocking buffer (5% skim milk, 1% fetal bovine serum, 0.2% tween 20 in PBS)...The capture reagent was His-tag Rabbit pAb (GenScript Cat. No. A00174). | Get A Quote |
| Plasmid DNA Preparation | Get A Quote | |
| Catalog Antibody | Get A Quote |
Vaccine induction of broadly neutralizing antibodies (bnAbs) to HIV remains a major challenge. Germline-targeting immunogens hold promise for initiating the induction of certain bnAb classes; yet for most bnAbs, a strong dependence on antibody heavy chain complementarity-determining region 3 (HCDR3) is a major barrier. Exploiting ultradeep human antibody sequencing data, we identified a diverse set of potential antibody precursors for a bnAb with dominant HCDR3 contacts. We then developed HIV envelope trimer-based immunogens that primed responses from rare bnAb-precursor B cells in a mouse model and bound a range of potential bnAb-precursor human naïve B cells in ex vivo screens. Our repertoire-guided germline... More
