کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
7608419 1493380 2018 10 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Preparation and evaluation of a hydrophilic interaction and cation-exchange chromatography stationary phase modified with 2-methacryloyloxyethyl phosphorylcholine
ترجمه فارسی عنوان
آماده سازی و ارزیابی یک تعامل هیدرولیکی و فاز ثابت کروماتوگرافی کاتیون مبادله اصلاح شده با 2-متاکریلویکسی اتیل فسفریکل کولین
کلمات کلیدی
2-متاکرییلویکسویید فسفوریل کولین، کروماتوگرافی متقابل هیدروفیلی، کروماتوگرافی تبادل کاتیونی ضعیف، ترکیبات قطبی، پروتئین،
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آنالیزی یا شیمی تجزیه
چکیده انگلیسی
In this work, 2-methacryloyloxyethyl phosphorylcholine (MPC) was used as a ligand to prepare a novel mixed-mode chromatography (MMC) stationary phase by the thiol-ene click reaction onto silica (MPC-silica). It was found that this MPC-silica showed the retention characteristics of hydrophilic interaction chromatography (HILIC) and weak cation exchange chromatography (WCX) under suitable mobile phase conditions. In detail, acidic and basic hydrophilic compounds and puerarin from pueraria were separated quickly with HILIC mode. Meanwhile, six standard proteins were allowed to reach baseline separation in WCX mode, and protein separation from egg white was also achieved with this mode. In addition, reduced/denatured lysozyme could be refolded with the MPC-silica column. In the meantime, the MPC-silica has been applied for refolding with simultaneous purification of recombinant human Delta-like1-RGD (rhDll1-RGD) expressed in Escherichia coli. The results show that the mass recovery and purity of rhDll1-RGD could reach 63.4% and 97% by one step, respectively. Furthermore, the reporter assay results demonstrated that refolded with simultaneously purified rhDll1-RGD could efficiently activate the signalling pathway in a dose-dependent manner. In general, this MPC-silica has good resolution and selectivity in the separation of polar compounds and protein samples in different high-performance liquid chromatography (HPLC) modes, and it successfully achieved refolding with simultaneous purification of denatured protein.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Chromatography A - Volume 1546, 20 April 2018, Pages 56-65
نویسندگان
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