کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1199696 1493599 2014 7 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Solid-phase polyethylene glycol conjugation using hydrophobic interaction chromatography
ترجمه فارسی عنوان
پیوند پلی اتیلن گلیکول فاز جامد با استفاده از کروماتوگرافی تعامل هیدروفوبی
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آنالیزی یا شیمی تجزیه
چکیده انگلیسی


• A novel solid-phase PEGylation using hydrophobic interaction chromatography (HIC).
• mPEG was first immobilized on the HIC, FGF-1 was then passed through the column.
• Lysozyme was first immobilized on the HIC, mPEG was then passed through the column.
• The modified rhFGF-1 and lysozyme retain good structural integrity and bioactivity.

PEGylation is a widely applied approach to improve the pharmacokinetic and pharmacodynamic properties of protein therapeutics. The current solution-phase PEGylation protocols often suffer from poor yield of homogeneously PEGylated bioactive products and hence fall short of being commercially attractive. To improve upon these techniques, here we developed a novel, solid-phase PEGylation methodology using a hydrophobic interaction chromatography (HIC) resin. Two variations of the HIC-based PEGylation are described that are tailored towards conjugation of proteins with hydrophobicity index above (lysozyme) and below (fibroblast growth factor 1, FGF-1) that of the mPEG-butyraldehyde (mPEG) chain used. In the case of lysozyme, the protein was first immobilized on the HIC, and the HIC-bound protein was then conjugated by passing over the column. In the case of FGF-1, the mPEG solution was first immobilized on the HIC, and the FGF-1 solution was then passed through the column. Circular dichroism (CD) spectroscopy demonstrated HIC-based PEGylation almost retained the secondary structures of proteins. Bioactivity assay showed that the recovery of activity of HIC-based PEGylated rhFGF-1 (i.e. 92%) was higher than that of liquid-phase PEGylated rhFGF-1 (i.e. 61%), while HIC-based PEGylated lysozyme showed the same activity recovery (i.e. 7%) as the liquid-phase PEGylated form. For specific proteins, the HIC-based solid-phase PEGylation maybe offer a more promising alternative than the current PEGylation methods and is expected to have a major impact in the area of protein-based therapeutics.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Chromatography A - Volume 1327, 31 January 2014, Pages 66–72
نویسندگان
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