Carbohydrate Microarrays Identify Blood Group Precursor Cryptic Epitopes as Potential Immunological Targets of Breast Cancer

Figure 2

Carbohydrate-specific ELISA and ELISA inhibition assays validate binding specificities of HAE3. (a) An antigen-specific ELISA distinguished HAE3 binding specificity from the T-antigen-specific PNA. ELISA plates were coated with O-core antigen, Tij II (31^#), T (81^#), and Tn (82^#) at 10 μg/mL to react with either HAE3 (5.0 μg/mL) or PNA (1.0 μg/mL). (b) ELISA inhibition assays with a series of carbohydrate antigens as competitors (25.0 μg/mL) to inhibit interaction between EPGN (1.0 μU/mL) and anti-HCA (HAE3, 1.0 μg/mL). These antigens include HCA (1^#), EPGN (85^#), which is the immunogen of HAE3, and blood group substances Tn (83^#), T (84^#), H (4^# or 22^#), A (3^#), B (14^#), Beach P1 (29^#), Tij II (31^#), and OG (32^#). Results are shown as percent inhibition in the presence of an inhibitor.

(a) ELISA binding assays

(b) ELISA inhibition assays