Purification of opioid receptors and preparation and application of monoclonal antibodies to them

阿片样物质受体被分为三种不同的亚型，g、δ和κ。阿片样物质受体被分为三种不同的亚型，μ、δ和κ。然而，阿片类药物通过这些不同亚型的药理作用机制的分子基础知之甚少。本文主要从阿片受体单克隆抗体的制备及应用两个方面来阐明阿片受体的分子机制。将神经母细胞瘤×神经胶质瘤杂交瘤NG 108 -15细胞膜免疫BALB/C小鼠4个月。将脾细胞与骨髓瘤细胞融合，并将含有杂交瘤的威尔斯孔中的培养上清液用于δ-阿片样物质结合实验中的测定。δ-OpR-1不仅表现出对δ-阿片样物质激动剂结合的有效抑制，其Bmax值降低，而且还表现出对δ-激动剂诱导的腺苷酸环化酶活性抑制的阻断。另一方面，δ-OpR-3显示出δ-阿片样物质激动剂结合的抑制，其亲和力降低。发现该抗体对NG 108 -15细胞中表达的δ-阿片受体进行免疫细胞化学染色。这些单克隆抗体将用于δ-阿片受体的纯化或其表达克隆的免疫筛选方法。

Opioid receptors are classified into three different subtypes, g,&, and K.Opioid receptors are classified into three different subtypes, mu,delta, and kappa. However, little is known of molecular basis of mechanisms of pharmacological actions of opioids through such different subtypes. The present works were focused on the preparation of monoclonal antibodies to opioid receptors and application in order to clarify such molecular mechanisms. Neuroblastoma x glioma hybrid NG108-15 cell membranes were immunized into BALB/C mice for 4 months. The spleen cells were fused with myeloma cells and the culture supernatants in wells containing hybridomas were used for assays in delta-opioid binding experiments. delta-OpR-l showed not only a potent inhibition of delta-opioid agonist binding with a decrease in its Bmax value, but also a blocking of delta-agonist-induced inhibition of adenylate cyclase activity. On the other hand, delta-OpR-3 showed an inhibition of delta-opioid agonist binding with a decrease in its affinity. This antibody was found to stain immunocytochemically delta-opioid receptor expressed in NG108-15 cells. These monoclonal antibodies will be useful for purification of delta-opioid receptors or its expression cloning by immunoscreening methods.