Foureau David M, Vrikkis Regina M, Jones Chase P, Weaver Katherine D, Macfarlane Douglas R, Salo Jonathan C, McKillop Iain H, Elliott Gloria D
Department of General Surgery, Carolinas Medical Center, Charlotte, NC.
Department of General Surgery, Carolinas Medical Center, Charlotte, NC ; Department of Mechanical Engineering and Engineering Science, UNC at Charlotte, Charlotte, NC.
Cell Mol Bioeng. 2012 Dec 1;5(4):390-401. doi: 10.1007/s12195-012-0243-x.
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. 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 T 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 T. 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 NaHCO (pH≥7.2) compared to controls without CDHP. While increased T is associated with a diminished rhIL-2 biological activity, the therapeutic protein remains structurally intact and functional.
磷酸二氢胆碱（CDHP）是一种离子液体，据报道它能提高模型蛋白的热稳定性。目前的研究考察了CDHP对重组人白细胞介素-2（rhIL-2）结构完整性和生物活性的影响，rhIL-2是一种用于治疗晚期黑色素瘤的治疗性蛋白。还使用长期暴露于该离子液体的原代细胞培养物或B16-F10细胞系评估了CDHP的生物相容性。与碱性缓冲液配方相比，在pH 7.4的680mM CDHP水溶液中配制rhIL-2，导致rhIL-2的熔点（Tm）升高了12.5°C，并且当溶液加热到高于熔点23.3°C时，能使rhIL-2构象稳定。CDHP溶液（≤80mM）对培养中的原代脾细胞或B16-F10细胞无细胞毒性活性。然而，与不含CDHP的对照相比，当rhIL-2用于含NaHCO₃（pH≥7.2）的30mM CDHP水溶液中时，观察到其生物活性损失了10倍。虽然熔点升高与rhIL-2生物活性降低有关，但该治疗性蛋白的结构仍保持完整且具有功能。
