Detailed characterization of reactivities of anti-gastric cancer monoclonal antibodies to carbohydrate antigen.

Four murine monoclonal antibodies (MoAbs), KM191, KM206, KM230 and KM231, raised against gastric cancer exhibited very similar reactivities to human carcinoma cells, whereas their abilities in probing the antigen shed from the cancer cells were quite different. We found in this study that the four MoAbs reacted immunologically with the same monosialo gangliosides derived from a gastric cancer cell line by two-dimensional high performance thin-layer chromatography. When the reactivities of the MoAbs to a number of purified gangliosides or oligosaccharides were examined, the carbohydrate structure of the antigen was determined as sialyl Lea. KM231 exhibited the highest binding avidity to the oligosaccharide and the ganglioside among the four MoAbs. In a comparative study of KM231 and NS19-9, which is a widely used sialyl Lea-reactive MoAb, KM231 bound to the oligosaccharide with higher affinity and detected the antigen in tissue sections with higher sensitivity. In addition, KM231 could detect a small amount of the antigen ganglioside in human gastric normal and cancerous mucosa and in gastric cancer cell lines by HPTLC-immunostaining.