Anti-mullerian hormone (AMH) is a cytokine of transforming growth factor β (TGF-β) superfamily able to induce apoptosis in cells bearing specific AMH type II receptors (AMHRII). AMHRII is overexpressed in some malignant cells, so at present recombinant AMH (rAMH) is considered as a new candidate antineoplastic drug. The use of rAMH may be especially effective in case of such severe diseases as ovarian, prostate and breast cancer. However, the development of a new drug is hampered by the laboriousness of obtaining highly purified rAMH and by the lack of data about the pharmacological characteristics of rAMH derivatives. In this work, we obtained preparations of prohormone, half-cleaved rAMH and a C-terminal fragment of rAMH, which was confirmed by qualitative and quantitative analyses. To obtain rAMH and its derivatives we used a previously developed highly effective producer strain containing the optimized human AMH gene. The production process has been divided into several stages: (a) rAMH biosynthesis in the bioreactor; (b) culture media preparation; (c) purification of rAMH and its derivatives using immunoaffinity chromatography and reversed-phase HPLC; (d) identification of the purified proteins by immunoblotting and analytical reversed-phase HPLC; and (e) evaluation of the hormone forms activity. The obtained proteins may be used in preclinical trials and in vitro study of rAMH derivatives properties.

中文翻译：

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人重组抗米勒激素及其衍生物的纯化。

抗苗勒激素（AMH）是转化生长因子β（TGF-β）超家族的细胞因子，能够诱导带有特定AMH II型受体（AMHRII）的细胞凋亡。AMHRII在某些恶性细胞中过表达，因此目前认为重组AMH（rAMH）是一种新的抗肿瘤药物。在诸如卵巢癌，前列腺癌和乳腺癌的严重疾病的情况下，使用rAMH可能特别有效。但是，由于难以获得高纯度的rAMH，以及缺乏有关rAMH衍生物的药理特性的数据，阻碍了新药的开发。在这项工作中，我们获得了激素，半切割的rAMH和rAMH的C端片段的制备方法，该方法已通过定性和定量分析得到证实。为了获得rAMH及其衍生物，我们使用了先前开发的，含有优化的人AMH基因的高效生产菌株。生产过程分为几个阶段：（a）生物反应器中rAMH的生物合成；（b）培养基的准备；（c）使用免疫亲和层析和反相HPLC纯化rAMH及其衍生物；（d）通过免疫印迹和分析型反相HPLC鉴定纯化的蛋白质；（e）评估激素形式的活性。获得的蛋白质可用于rAMH衍生物特性的临床前试验和体外研究。（c）使用免疫亲和层析和反相HPLC纯化rAMH及其衍生物；（d）通过免疫印迹和分析型反相HPLC鉴定纯化的蛋白质；（e）评估激素形式的活性。获得的蛋白质可用于rAMH衍生物特性的临床前试验和体外研究。（c）使用免疫亲和层析和反相HPLC纯化rAMH及其衍生物；（d）通过免疫印迹和分析型反相HPLC鉴定纯化的蛋白质；（e）评估激素形式的活性。获得的蛋白质可用于rAMH衍生物特性的临床前试验和体外研究。