Fig. 3.

Kinetics of humoral immune response elicited by Galα3LN-HSA, Galα3LN-HSA+LMPLA, and HSA+LMPLA. Chemiluminescent enzyme-linked immunosorbent assay (CL-ELISA) reactivity of mouse sera from α1,3GalT-KO mice injected with Galα3LN-HSA, Galα3LN-HSA+LMPLA, or HSA+LMPLA using Galα3LN-HSA as immobilized antigen. For immunoglobulin M (IgM) measurement, prior to the CL-ELISA we depleted IgG antibodies by preincubating each serum sample with a 15-μL bead suspension mixture of protein A-Sepharose 4B and protein G-Sepharose, both fast flow (A:G, 1:1, v/v), and recombinant human serum albumin (HSA; 1 mg/mL), for 1 h at 37 °C on rotatory shaker. Error bars indicate S.E.M. of triplicate determinations. Sample no longer available is indicated (#). RLU relative luminescence units