Silkworm-Derived GM1 Affinity Assay

A GM1-ELISA was performed as described previously [13 (link)] to detect the affinity of silkworm-derived fusion proteins for GM1 ganglioside. The microtiter plates were coated with monosialoganglioside-GM1 (Sigma, USA) by incubating the plates with 50 μl/well GM1 in methanol at 4°C overnight. The wells were then blocked with BSA solution, and the hemolymph dilutions were added. The same dilutions of normal hemolymph and serial dilutions of bacterial CTB were used as a negative control and to generate the standard curve, respectively. The remainder of the procedure was identical to the semiquantitative ELISA assay described above.