KAPLAN HOSP,REHOVOT,ISRAEL. HEBREW UNIV JERUSALEM,FAC AGR,DEPT BIOCHEM FOOD SCI & NUTR,IL-76100 REHOVOT,ISRAEL. LANIADO HOSP,NETANYA,ISRAEL.
Oncol Rep. 1995 Jul;2(4):679-83. doi: 10.3892/or.2.4.679.
A new method for the isolation of p53 protein from the sera of cancer and non-cancer patients has been developed. The method is based on the utilization of a new modification of a support for affinity chromatography in the form of gel fiberglass (GFG) (R. Zusman, patent applications, Israel, 1992; USA 1993). The sera were percolated through GFG columns with entrapped rabbit IgG generated against the colon cancer antigens. Tumor-associated antigens were eluted from sera of cancer patients in large amounts, up to 2.4 mg/ml serum/column. Concentrations of these antigens in different sera and their affinity to anti-tumoral antibodies were detected by ELISA. Western immunoblotting with commercial monoclonal antibodies and SDS-PAGE identified the isolated antigens as the p53 protein. This protein has been isolated in the highest concentration yet reported from the serum of cancer patients, up to 1 mg/ml. The same protein has been found in the serum of healthy people and non-cancer patients but in significantly lower concentrations. Isolation of p53 protein from sera of noncancer patients confirms the opinion of some investigators that this protein should be considered not only as an oncogene but also as a protein correlated with different changes in the cell cycle.
已开发出一种从癌症和非癌症患者的血清中分离 p53 蛋白的新方法。该方法基于利用亲和层析支持物的新修饰形式，即玻璃纤维凝胶（GFG）（R. Zusman，专利申请，以色列，1992 年；美国，1993 年）。将血清通过与结肠癌抗原结合的兔 IgG 包埋的 GFG 柱进行渗滤。从癌症患者的血清中大量洗脱肿瘤相关抗原，高达 2.4mg/ml 血清/柱。通过 ELISA 检测不同血清中这些抗原的浓度及其与抗肿瘤抗体的亲和力。用商业单克隆抗体进行 Western 免疫印迹和 SDS-PAGE 鉴定分离的抗原为 p53 蛋白。这种蛋白质已从癌症患者的血清中分离出来，浓度高达 1mg/ml，这是迄今为止报道的最高浓度。在健康人和非癌症患者的血清中也发现了相同的蛋白质，但浓度明显较低。从非癌症患者的血清中分离 p53 蛋白证实了一些研究人员的观点，即该蛋白不仅应被视为癌基因，还应被视为与细胞周期中不同变化相关的蛋白质。
