The human androgen receptor (hAR) is an important regulatory protein particularly in male sexual differentiation. The investigation of hAR functionality has been hampered by the lack of AR specific monoclonal antibodies recognizing the functional domains of the receptor. Therefore production of high affinity mono-specific polyclonal (PAbs) and monoclonal antibodies (MAbs) directed to the hAR was initiated following the synthetic peptide (SP) strategy. Five hAR specific peptides were selected on the basis of their predicted antigenic properties avoiding homology with other steroid hormone receptors. Peptide specific polyclonal antisera were obtained following selected immunization protocols. Mono-specific polyclonal antibody responses were elicited to all peptides in mice and rabbits. Crossreactivity of the peptide specific antisera with the native hAR in various biochemical assays was observed with two out of five peptides. Peptide SP61 (hAR residues 301-320) was used for the generation site-directed MAbs specific for the hAR. Specificity for the hAR was established by immunoprecipitation, immune-complex density gradient centrifugation and immunohistochemistry on human prostate tissue sections. The multi-assay performance of the selected high affinity antibodies proved the usefulness of the straight forward peptide approach and opens a wide field of possible biochemical and physiological investigations into questions related to androgen action.

译文

人类雄激素受体 (hAR) 是一种重要的调节蛋白,尤其是在男性性别分化中。缺乏识别受体功能域的AR特异性单克隆抗体,阻碍了对hAR功能的研究。因此,按照合成肽 (SP) 策略开始生产针对hAR的高亲和力单特异性多克隆 (PAbs) 和单克隆抗体 (mab)。根据其预测的抗原特性选择了五种hAR特异性肽,以避免与其他类固醇激素受体的同源性。按照选定预防接种方案获得了肽特异性多克隆抗血清。对小鼠和兔子的所有肽均产生单特异性多克隆抗体反应。在各种生化测定中,用五分之二的肽观察到肽特异性抗血清与天然hAR的交叉反应性。肽SP61 (hAR残基301-320) 用于hAR特异性的生成定点mab。通过免疫沉淀,免疫复合物密度梯度离心和免疫组织化学对人前列腺组织切片确定hAR的特异性。所选的高亲和力抗体的多测定性能证明了直接肽方法的有用性,并为与雄激素作用有关的问题开辟了可能的生化和生理研究的广阔领域。

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