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目的 用欧蔓陀罗凝集素 (DSA)分离血清γ 谷氨酰转移酶 (GGT)糖链的强结合组份 ,观察正常人、良性肝病及原发性肝癌 (PHC)患者的变化 ,为建立小柱亲和层析法分离PHC特异性GGT糖链奠定基础。方法 以神经胺酸酶 (NMD)水解后的血清直接上DSA亲和层析柱 ,首先用0 0 5M醋酸洗去不结合和弱结合组份 ,再用 0 1M醋酸洗脱强结合组份 ,用连续监测法测定洗脱液的GGT活性变化。结果 正常人、良性肝病患者在 0 0 5M醋酸洗脱时可出现 1～ 2个峰 ,没有强结合峰。PHC除此之外还出现一个比例不大的强结合峰。结论 采用二步法洗脱DSA亲和层析柱 ,可以较好地分离出DSA强结合组份。该组份为PHC患者所特有 ,PHC患者有较高的阳性率(80 % )。
OBJECTIVE: To isolate the strong binding components of GGT by DSA, and to observe the changes of normal, benign liver disease and primary hepatocellular carcinoma (PHC) Column affinity chromatography method to isolate PHC-specific GGT sugar lays the foundation. Methods After the serum hydrolyzed by neuraminidase (NMD) was directly on DSA affinity chromatography column, the unbound and weakly bound components were washed away with 0 0 5 M acetic acid first, then the strong bound components were eluted with 0 1M acetic acid, Continuous monitoring method was used to determine the change of GGT activity of the eluate. Results In normal subjects, patients with benign liver disease showed 1-2 peaks when eluted with 0 0 5 M acetic acid and no strong binding peak. PHC in addition there is a small proportion of the strong combination of peaks. Conclusion DSA affinity chromatography was eluted by two-step method and the strong binding component of DSA could be separated well. This component is specific to patients with PHC, PHC patients have a higher positive rate (80%).