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. 2019 Jun 4;10:1254. doi: 10.3389/fimmu.2019.01254

Figure 4.

Figure 4

Functional activity of anti-S-antigen and anti-AARP antibodies in the assay of GIA. (A) Titration of the same anti-AARP-BAP-Strep sample shown in Figure 3B against the 3D7 clone and FVO strain. (B) Effect of RBC pre-incubation on a range of mouse antibody specificities. TRA-1-85 is a mouse anti-basigin monoclonal antibody. Other samples are total IgG from immunized mice (see Figure S3 for further details). (C) Pre-mixing of the relevant immunizing antigen with mouse total IgG. Final antigen concentration was 1.35 mg/mL, IgG concentration was 1 mg/mL. (D) Titration of the same anti-S-antigen-BAP-Strep sample shown in Figure 3B against the 3D7 clone and FVO strain. (E) Total IgG from two Zika rabbits immunized with S-antigen-BAP-Strep formulated with Freund's adjuvant system. Samples were subjected to pre-depletion with RBCs. (A–D) are each from a single GIA assay run due to sample volume constraints. (A,D), lines are connected mean and points individual replicate wells (two per concentration). (F) Pre-mixing of S-antigen protein with pooled rabbit total IgG from (E). Final S-antigen concentration was 1.5 mg/mL, total IgG concentration was 10 mg/mL. (A–D), mean and range of four replicates. (E,F), mean and range of three wells of a single GIA assay, representative of data from two replicate GIA assay runs.