人胎肝金属硫蛋白的分离纯化及酶联免疫吸附检测

Purification and Enzyme Linked Immunosorbent Assay of Human Fetal Liver Metallothionein

人胎肝经匀浆、离心、乙醇沉淀后，通过ＳｅｐｈａｄｅｘＧ－７５、ＤＥＡＥ－ＳｅｐｈａｒｏｓｅＦａｓｔＦｌｏｗ柱层析分离，获得金属硫蛋白的两种亚型（ＭＴ－１和ＭＴ－２）。经氨基酸组成分析证明是均一的。用ＭＴ－１为抗原免疫青紫兰兔制备了抗血清，以纯化的ＩｇＧ建立了人ＭＴ的酶联免疫吸附检测（ＥＬＩＳＡ）方法，并给出了标准工作曲线。

After human fetal liver was homogenized,centrifuged and precipitated with ethanol,two forms of metallothionein (MT 1,MT 2) were purified by gel filtration chromatography on Sephadex G 75,ion exchange chromatography on DEAE Sepharose Fast Flow.Their purity was verified by amino acid composition analysis.Using human MT 1 as antigen,the antiserum was produced in rabbit.Then enzyme linked immunosorbent assay (ELISA) was developed for the detection and quantitative analysis of human metallothionein,and the standard curves were supplied.