Experimental study on intraoperative diagnosis of metastatic region

亚细胞分裂法检测82k/6.3和61k/7.5的定位。在与7种凝集素的结合反应中，82K/6.3与Rca I、DBA和WGA反应，61K/7.5与Rca I、DBA、WGA、UEA I和SBA结合。两种蛋白免疫的BALB/C小鼠的脾细胞与小鼠骨髓瘤细胞融合，可产生抗82K/6.3或61K/7.5的单抗。在间接免疫荧光研究中，抗82K/6.3和61K/7.5的单抗分别与人癌细胞的胞浆和细胞膜反应。这一结果有力地提示了亚细胞分裂证明这两种蛋白的定位。给荷瘤裸鼠静脉注射同位素标记的单抗后，在第一天、第三天、第五天拍摄免疫核素图。由于碘不能阻断甲状腺，大量的碘蓄积了甲状腺。然而，右侧大腿种植的肿瘤也显示出大量的碘蓄积。肿瘤/心血比值第1天为1.46，第3天为1.88-0.21，第5天为2.36-0.89。这一结果有望应用于临床研究。

The localization of 82k/6.3 and 61k/7.5 was investigated by subcellular fractionation. They were rich in Microsomal fraction, but not found in both nuclear and mitochondrial fractions.In binding reaction with seven kinds of lectins, 82k/6.3 reacted with RCA I, DBA and WGA, where 61k/7.5 reacted with RCA I, DBA,WGA,UEA I and SBA. Transferrin:, reacted with only RCA I.Each hybrid producing monoclonal antibody against 82k/6.3 or 61k/7.5 was generated by fusing spleen cell of BALB/C mice immunized by the two proteins and mouse myeloma cells. Each monoclonal antibody was specified in enzyme-linked immunoassay.In indirect immune-fluorescent studies, monoclonal antibodies against 82k/6.3 and 61k/7.5 reacted with cytoplasma and membrane of human cancer cell. This result strongly suggests the localization of the two proteins demonstrated by subcellular fractionation.After iv administration of isotope-labelled monoclonal antibody to tumor- bearing nude mouse, immunoscintigram was taken on i, 3,5 day. Because of no block by iodine, thyroid was accumulated by large amount of iodine. However, implanted tumor at right thigh showed also large amount of accumulation of iodine. Ratio of Tumor/heart blood was 1.46 in 1st day, 1.88<plus-minus>0.21 on 3rd day and 2.36<plus-minus>0.89 on 5th day. This result was promising to apply to clinical investigat