MOLECULAR ASPECTS OF THE MULTIPLE OPIOID RECEPTORS

该建议针对产生和表征单克隆抗体。 多种阿片受体的抗体 反过来，这些抗体 将被用于表征和纯化阿片受体。 单克隆 受体的抗体将通过放射免疫测定法筛选， 免疫沉淀和它们抑制阿片结合的能力。 一 将更详细地研究已经产生的抗体OR-465.3。 研究将开始，以确定哪种类型的阿片受体，这 抗体是针对。 针对本发明的其它单克隆抗体 受体将通过使用大鼠神经膜产生，溶解 阿片受体和神经母细胞瘤-胶质瘤，NG 108 -15，膜作为抗原。 针对每种阿片受体的特异性抗体是最终目标。 将测试抗体抑制阿片剂结合至 大鼠神经膜和溶解的受体制剂。 他们的 对阿片剂与外周组织如豚鼠回肠结合的影响 并检查小鼠输精管。 免疫沉淀法 来自溶解制剂的受体将用于鉴定抗体 与受体相互作用，但不在阿片结合位点。 将通过蛋白质印迹技术检测抗体，以确定 蛋白质（S），他们针对。 最后，这些抗体将 用于纯化它们识别的阿片类药物结合蛋白。 通过使用 单克隆抗体，将有可能探测阿片受体（S） 直接.

This proposal is directed at generating and characterizing monoclonal antibodies to the multiple opioid receptors. In turn, these antibodies will be used to characterize and purify the opioid receptors. Monoclonal antibodies to the receptor will be screened by radioimmunoassay, immunoprecipitation and on their ability to inhibit opiate binding. One antibody already generated, OR-465.3, will be studied in greater detail. Studies will be initiated to ascertain which type of opioid receptor, this antibody is directed against. Additional monoclonal antibodies to the receptor will be generated by using rat neural membranes, solubilized opioid receptor and neuroblastoma-glioma, NG108-15, membranes as antigens. Antibodies specific for each type of opioid receptor is the ultimate goal. Antibodies will be tested for their ability to inhibit opiate binding to rat neural membranes and to solubilized receptor preparations. Their effect on opiate binding to peripheral tissues such as the guinea pig ileum and the mouse vas deferens will be examined. Immunoprecipitation of the receptor from a solubilized preparation will be used to identify antibodies that interact with the receptor but, not at its opiate binding site. Antibodies will be tested by a Western blot technique to ascertain which protein(s) they're directed against. Finally, these antibodies will be used to purify the opiate binding protein(s) that they recognize. By using monoclonal antibodies, it will be possible to probe the opioid receptor (s) directly.