Diagnostic method for differentiation of hepatocellular carcinoma : Determination of immature TGFα in blood and urine.

protef - α经过加工成为tgf - α的成熟形式。在这项研究中，我们合成了几种氨基酸序列仅存在于proTGFα序列中而不存在于成熟形式TGFα序列中的肽。用其中一种肽免疫新西兰大白兔。我们利用抗血清的免疫球蛋白G部分和抗TGFα的多克隆抗体开发了夹心ELISA。取正常成人或肝细胞癌患者尿液，加入终浓度为70%的硫酸铵离心。沉淀馏分作为测定样品。新开发的ELISA不能检测成熟形态的tgf - α。然而，它检测到TGFα的未成熟形式，其分子量大于成熟形式。在肝细胞癌患者的尿液中检测到未成熟形态的tgf - α，而在正常成人的尿液中未检测到。从这些结果来看，tgf - α的未成熟形式可能在肝细胞癌患者中升高。测定未成熟形态的TGFα可用于肝细胞癌的诊断。我们现在正在分析肝细胞癌患者血液中未成熟形态TGFα的含量。

ProTGFα is processed to become a mature form of TGFα. In this study, we synthesized several kinds of peptides, amino acid sequences of which exist only in the sequence of proTGFα but not in that of the mature form of TGFα. New Zealand white rabbits were immunized with one of the peptides. We developed a sandwich ELISA using immunoglobulin G fraction of the antiserum and polyclonal antibodies against TGFα. Urine collected from normal adults or patients with hepatocellular carcinoma was added with ammonium sulfate at the final concentration of 70 percent and centrifuged. Precipitate fraction was used as an assay sample. The newly developed ELISA did not detect the mature form of TGFα. However, it detected an immature form of TGFα, the molecular weight of which was larger than that of the mature form. The immature form of TGFα was detected in urine from patients with hepatocellular carcinoma but not in that from normal adults. From these results, it is possible that the immature form of TGFα is elevated in patients with hepatocellular carcinoma. Determination of the immature form of TGFα can be useful for diagnosis of hepatocellular carcinoma. We are now analyzing the content of the immature form of TGFα in blood from patients with hepatocellular carcinoma.