Interleukin-2 mutants with enhanced alpha-receptor subunit binding affinity.

Protein engineering Pub Date : 2003-12-01 DOI:10.1093/protein/gzg111

Balaji M Rao, Andrew T Girvin, Thomas Ciardelli, Douglas A Lauffenburger, K Dane Wittrup

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引用次数: 54

Abstract

Stimulation of T-cells by IL-2 has been exploited for treatment of metastatic renal carcinoma and melanoma. However, a narrow therapeutic window delimited by negligible stimulation of T-cells at low picomolar concentrations and undesirable stimulation of NK cells at nanomolar concentrations hampers IL-2-based therapies. We hypothesized that increasing the affinity of IL-2 for IL-2Ralpha may create a class of IL-2 mutants with increased biological potency as compared with wild-type IL-2. Towards this end, we have screened libraries of mutated IL-2 displayed on the surface of yeast and isolated mutants with a 15-30-fold improved affinity for the IL-2Ralpha subunit. These mutants do not exhibit appreciably altered bioactivity at 0.5-5 pM in steady-state bioassays, concentrations well below the IL-2Ralpha equilibrium binding constant for both the mutant and wild-type IL-2. A mutant was serendipitously identified that exhibited somewhat improved potency, perhaps via altered endocytic trafficking mechanisms described previously.

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具有增强α受体亚单位结合亲和力的白介素-2突变体。

IL-2刺激t细胞已被用于转移性肾癌和黑色素瘤的治疗。然而，在低皮摩尔浓度下对t细胞的可忽略刺激和在纳摩尔浓度下对NK细胞的不良刺激所限定的狭窄治疗窗口阻碍了基于il -2的治疗。我们假设，增加IL-2对il - 2rα的亲和力可能会产生一类与野生型IL-2相比具有更高生物效力的IL-2突变体。为此，我们筛选了显示在酵母表面的突变IL-2文库和分离的突变体，这些突变体对il - 2rα亚基的亲和力提高了15-30倍。在稳态生物测定中，这些突变体在0.5- 5pm时没有表现出明显的生物活性变化，浓度远低于突变体和野生型IL-2的il - 2rα平衡结合常数。偶然发现了一种突变体，其效力有所提高，可能是通过改变先前描述的内胞运输机制。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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Protein engineering

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