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The co-expression of denileukin diftitox immunotoxin with Artemin: soluble and aggregation analysis in presence of an efficient protein chaperone
Biologia ( IF 1.5 ) Pub Date : 2021-08-05 , DOI: 10.1007/s11756-021-00846-2
Mohamad Najarasl 1 , Mehdi Zeinoddini 1 , Ali Reza Saeeidinia 1 , Reza Hasan Sajedi 2
Affiliation  

Denileukin diftitox is the first Food and Drug Administration (FDA) approved recombinant immunotoxin that has been prescribed in the treatment of some leukemia and lymphoma such as cutaneous T-cell lymphoma, chronic lymphocytic leukemia, and B-cell non-Hodgkin’s lymphoma in which the IL-2 receptor is overexpressed. This smart and directed toxin was named DT389IL-2 because it contains truncated diphtheria toxin (DT387), which is fused to human interleukin 2 (IL2). Today, since this protein is not stable for a long time, its production is dependent on customer demand. Aggregation is an important obstacle in the recombinant production of heterologous proteins, especially those with therapeutic properties. This study aimed to synthesize denileukin diftitox in the presence of Artemin as an efficient chaperone in the E. coli expression system, as well as to assess the soluble/aggregation of the synthesized protein. Two plasmids, one containing the denileukin diftitox protein-coding gene and the other carrying artemin gene were simultaneously transferred into E. coli BL21 (DE3). The transformed bacteria were then induced to overexpress the mentioned proteins. The soluble expression of the proteins was confirmed by the native PAGE technique. The results confirmed that the soluble expression of the denileukin diftitox protein significantly increased in the presence of Artemin as a new chaperon protein. To conclude, this strategy can be utilized for the soluble expression and production of this protein as a smart and directed toxin for the treatment of recurrent cutaneous T-cell lymphoma (CTCL).



中文翻译:

Denileukin diftitox 免疫毒素与 Artemin 的共表达:在有效蛋白质伴侣存在下的可溶性和聚集分析

Denileukin diftitox是美国食品和药物管理局( FDA) 批准的首个重组免疫毒素,用于治疗某些白血病和淋巴瘤,如皮肤 T 细胞淋巴瘤、慢性淋巴细胞白血病和 B 细胞非霍奇金淋巴瘤,其中IL-2 受体过度表达。这种智能且定向的毒素被命名为 DT 389 IL-2,因为它含有与人白介素 2 (IL2) 融合的截短白喉毒素 (DT387)。今天,由于这种蛋白质长时间不稳定,其生产取决于客户的需求。聚集是异源蛋白质重组生产的一个重要障碍,尤其是那些具有治疗特性的蛋白质。本研究旨在合成denileukin diftitox在存在 Artemin 作为大肠杆菌表达系统中的有效伴侣的情况下,以及评估合成蛋白质的可溶性/聚集。两个质粒,一个含有denileukin diftitox蛋白编码基因,另一个携带artemin基因,同时转移到大肠杆菌BL21(DE3)中。然后诱导转化的细菌过度表达提到的蛋白质。蛋白质的可溶性表达由天然 PAGE 技术证实。结果证实denileukin diftitox的可溶性表达在 Artemin 作为一种新的伴侣蛋白存在的情况下,蛋白质显着增加。总而言之,该策略可用于该蛋白质的可溶性表达和生产,作为治疗复发性皮肤 T 细胞淋巴瘤 (CTCL) 的智能和定向毒素。

更新日期:2021-08-10
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