In vitro assessment of choline dihydrogen phosphate (CDHP) as a vehicle for recombinant human interleukin-2 (rhIL-2).
In vitro assessment of choline dihydrogen phosphate (CDHP) as a vehicle for recombinant human interleukin-2 (rhIL-2).
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DOI：
10.1007/s12195-012-0243-x
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发表时间：
2012-12-01
影响因子：
2.8
通讯作者：
Elliott, Gloria D.
中科院分区：
文献类型：
--
作者：
Foureau, David M.;Vrikkis, Regina M.;Jones, Chase P.;Weaver, Katherine D.;MacFarlane, Douglas R.;Salo, Jonathan C.;McKillop, Iain H.;Elliott, Gloria D.
磷酸胆碱二氢盐(CDHP)是一种离子液体，据报道可提高模型蛋白质的热稳定性。本工作研究了CDHP对重组人白介素2(rhIL-2)结构完整性和生物活性的影响，重组人白介素2是一种用于治疗晚期黑色素瘤的治疗蛋白。在体外，CDHP的生物相容性也通过长期暴露在离子液体中的原代细胞培养或B16-F10细胞株进行评估。在pH 7.4的680 mM CDHP水溶液中配制rhIL-2，与基本缓冲液配方相比，rhIL-2的Tm增加12.5°C，并在溶液加热到Tm以上23.3°C时提供构象稳定。CDHP液(≤80 mM)对原代脾细胞和B16-F10细胞无细胞毒作用。然而，在含有NaHCO3(pH≥7.2)的30 mM CDHP水溶液中，观察到重组人IL-2的生物活性是不含CDHP的对照组的10倍。虽然TM增加与rhIL-2生物活性降低有关，但治疗性蛋白保持结构完整和功能不变。
Choline dihydrogen phosphate (CDHP) is an ionic liquid reported to increase thermal stability of model proteins. The current work investigated CDHP effect on structural integrity and biological activity of recombinant human interleukin-2 (rhIL-2), a therapeutic protein used for treating advanced melanoma. In vitro CDHP biocompatibility was also evaluated using primary cell cultures, or B16-F10 cell line, chronically exposed to the ionic liquid. Formulation of rhIL-2 in an aqueous 680mM CDHP pH 7.4 solution resulted in a 12.5°C increase in the Tm of rhIL-2 compared to a basic buffer formulation, and provided conformational rhIL-2 stabilization when the solution was heated to 23.3°C above the Tm. CDHP solutions (≤80mM), exhibited no cytotoxic activity toward primary splenocytes or B16-F10 cells in culture. However, a 10-fold loss in biological activity was observed when rhIL-2 was used in a 30mM CDHP aqueous solution with NaHCO3 (pH≥7.2) compared to controls without CDHP. While increased Tm is associated with a diminished rhIL-2 biological activity, the therapeutic protein remains structurally intact and functional.
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影响因子：
2.9
作者：
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通讯作者：
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