Facile purification of mono-PEGylated interleukin-1 receptor antagonist and its characterization with multi-angle laser light scattering |
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| Institution: | 1. National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100080, PR China;2. The Graduate University of the Chinese Academy of Sciences, Beijing 100039, PR China;3. Beijing Baiao Pharmaceuticals Co., Ltd., Beijing 102200, PR China;1. Research Center of Siyuan Natural Pharmacy and Biotoxicology, College of Life Sciences Zhejiang University, Hangzhou 310058, China;2. Department of Pharmaceutical Analysis and Drug Metabolism, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, Zhejiang Province 310058, China;1. Departamento Química Analítica, Facultad de Química, Universidad de Valencia, Dr. Moliner 50, Burjassot, 46100 Valencia, Spain;2. Departamento Química Orgánica, Facultad de Química, Universidad de Valencia, Dr. Moliner 50, Burjassot, 46100 Valencia, Spain;1. Key Laboratory of Tibetan Medicine Research, Northwest Institute of Plateau Biology, Chinese Academy of Sciences, Xining 810001, China;2. Graduate University of the Chinese Academy of Sciences, Beijing 100049, China;1. SEMM, European School of Molecular Medicine, Campus IFOM-IEO, Via Adamello 16, 20139 Milano, Italy;2. Fondazione Filarete, Viale Ortles 22/4, 20139 Milano, Italy;3. CIMaINa, Dipartimento di Fisica, Università degli Studi di Milano, Via Celoria 16, 20133 Milano, Italy |
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| Abstract: | Recombinant human interleukin-1 receptor antagonist (rhIL-1ra) was chemically conjugated with succinimidyl carbonate monomethoxyl polyethylene glycols of 5 kDa (SC-PEG5k) and 10 kDa (SC-PEG10k) molecular weight. A facile purification of the conjugates was achieved by one-step cationic exchange chromatography. The purity of mono-PEGylated protein was greater than 95%. The purified conjugate was characterized by multi-angle laser light scattering (MALLS) for determining the apparent gyration radius (rg) and hydrodynamic radius (rh). MALLS results showed that the conjugation of PEG markedly enhanced rg and rh of parent protein (rg: from 15.7 to 48.2 nm for the PEG5k and 81.9 nm for the PEG10k; rh: from 4.2 to 58.4 nm for the PEG5k and 102.3 nm for the PEG10k). The PEGylated rhIL-1ra retained 44.6% of binding activities to the cell receptor for PEG5k and 40.2% for PEG10k, compared to the original protein. |
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