Fig. 1: DANA in vitro assessment of Env trimer antigenicity.
From: Assessing immunogenicity barriers of the HIV-1 envelope trimer

a Workflow of DANA. b DANA 1–5: Overview of recombinant HIV-1 Env trimer used for panning (I) and type and number of ribosome display selection rounds (II). c and d DARPin clones with valid ORF derived from the randomly picked 190 clones in each DANA screen were analyzed for binding and neutralizing properties (n = 126–184). Data are derived from single experiments. c Binding properties of DARPins obtained from DANA 1–5, based on ELISA, using Env trimer probes and the V3-crown mimetic peptide V3-IF (BG505) as specified in Supplementary Table 3. Trimer binding is categorized as binding to at least one of several trimers but not the V3-mimetic. V3 specificity includes solely V3-reactive and V3- and trimer dual-reactive clones. Low-level binders categorize DARPins with no trimer- and no V3-reactivity >3-fold over background. d Neutralization capacities of DARPins derived in DANA 1–5 against a multi-clade 5-pseudovirus panel according to a neutralization score reflecting breadth and potency. The max. score is 75, a score 5–14 indicates low neutralizing activity, a score of >15 indicates high neutralizing activity. The left panel depicts the neutralization score, the right panel the number of clones with low and high neutralizing activity, respectively.