de Groot P G, Hamers M N, Westerveld A, Schram A W, Meera Khan P, Tager J M
Hum Genet. 1978 Nov 16;44(3):295-304. doi: 10.1007/BF00394294.
An immunochemical method has been developed for the quantitative determination of species-specific gene products, for instance alpha-galactosidase and N-acetyl-alpha-galactosaminidase, in man-rodent hybrid cells and in the parental cell lines. Antisera raised against the purified enzymes are covalently coupled to Sepharose 4B. The gene products are specifically removed from a cell lysate by incubating with the appropriate Sepharose-coupled antiserum. After centrifugation followed by washing of the precipitated Sepharose, the enzymic activity can be quantitatively measured on the Sepharose beads. With this technique it has been demonstrated that the ability of human N-acetyl-alpha-galactosaminidase (also known as alpha-galactosidase B) to hydrolyze alpha-galactosidic linkages is lost when the enzyme is expressed in man-Chinese hamster hybrid cells.
已开发出一种免疫化学方法,用于定量测定人-鼠杂交细胞及其亲本细胞系中物种特异性基因产物,例如α-半乳糖苷酶和N-乙酰-α-半乳糖胺酶。针对纯化酶产生的抗血清与琼脂糖4B共价偶联。通过与适当的琼脂糖偶联抗血清孵育,可从细胞裂解物中特异性去除基因产物。离心后洗涤沉淀的琼脂糖,然后可在琼脂糖珠上定量测量酶活性。通过该技术已证明,当人N-乙酰-α-半乳糖胺酶(也称为α-半乳糖苷酶B)在人-中国仓鼠杂交细胞中表达时,其水解α-半乳糖苷键的能力丧失。