Journal of Immunology Research / 2015 / Article / Fig 2

Research Article

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

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