Detection of prolactin based on the receptor-binding activity and its application

催乳素通过与其位于质膜上的受体结合，向靶器官传递荷尔蒙信息。Shill等人。有报道称，催乳素的受体结合活性与其生物学活性有很好的相关性。-然而，催乳素的各种分子形式已经被确定。其中，糖基化催乳素的活性低于非糖基化催乳素。微粒体组分已被用作受体的来源。在本实验中，我们用Triton X-100溶解这些膜蛋白，用Con-A亲和层析进行部分纯化，然后将它们固定在硝酸纤维素膜过滤器上。用膜结合型受体和牛血清，用放射受体测定法测定血清催乳素水平。测定范围为0.4~20 ng/ml，血清浓度最高可达测定缓冲液的16%，检出限为2.5 ng/ml。两种方法的比值接近0.99(r&gt；+0.99，n=55)。两种方法的凝胶过滤图谱是一致的。结论膜结合受体可用于血清催乳素的测定，结果令人满意。

Prolactin sends its hormonal message to target organs through binding with its receptor located on the plasma membrane. Shill et al. reported that the receptor-binding activity of prolactin correlates well with its biological activity. -However, various molecular forms of prolactin have been identified. Among them, for example glycosylatede prolactin is less active than nonglycosylated prolactin. The microsome fraction has been used as a source of the receptor. In the present experiments, we solubilized these membrane proteins with Triton X-100, purified partially using Con-A affinity chromatography, and then immobilized them on a nitrocellulose membrane filter. Using membrane-bound receptor and bovine serum, the serum level of prolactin was determined by radioreceptor assay. The dosedisplacement curve was obtained in the range of 0.4 and 20 ng/ml. Serum could be included in concentrations up to 16% of the assay buffer and, thus the detection limit was about 2.5 ng/ml. Prolactin in the serum was determined by mdioimmunoassay. The ratio of the two assays was nearly 0.99 (r>+O. 99, n=55). The gel filtration profile was identical between the two assays. It was concluded that membranebound receptor can be used for the determination of serum prolactin with satisfactory.