ISOLATION AND ANALYSIS OF LECTIN-REACTIVE SARCOMA-ASSOCIATED MEMBRANE-GLYCOPROTEINS

Sarcoma and normal tissue plasma membrane lectin-reactive glycoprotein s were analyzed by sodium dodecyl sulfate polyacrylamide gel electroph oresis. Two peanut agglutinin-reactive N-acetylgalactosamine-containin g glycoproteins of 1.05x10(6) and 1.25x10(5) Da and one lentil aggluti nin-reactive mannose/N-acetylglucosamine (-fucose)/sialic acid-contain ing glycoprotein of 1.7x10(5) Da (Gp170) were detected in osteosarcoma and malignant fibrous histiocyctoma (MFH), respectively. However, the se glycoproteins were not detected in normal tissue plasma membranes. Concanavalin, A, wheat germ and Ulex europaeus Type I agglutinins did not reveal any unique sarcoma-associated membrane glycoproteins. Preli minary studies on monoclonal antibodies (mAbs) generated against Gp170 (mAb 64-35-84) and against lentil-reactive glycoproteins from MFH (mA bs 67-34 and 67-117) revealed high specific binding to a number of mem branes isolated from MFH and ostoesarcoma tissues, with no crossreacti vity to normal human tissues tested (liver, spleen and skin). Detailed analysis of mAb 67-102, which was generated against lentil-reactive g lycoproteins isolated from MFH plasma membranes, exhibited significant binding to membranes isolated from osteosarcoma, liposarcoma and MFH; moderate binding to synovial sarcoma, aggressive fibromatosis and fib rosarcoma; and minimal to no binding to other soft tissue sarcoma plas ma membranes. No binding was observed to twenty normal tissue specimen s, with the exception of low positive binding to two of five fat and t wo of three colon specimens.