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中国生物工程杂志

China Biotechnology
China Biotechnology  2015, Vol. 35 Issue (12): 109-114    DOI: 10.13523/j.cb.20151217
    
Quality Control for PEGylated Recombinant Protein
LI Xing1,2, YAO Wen-bing1, XU Chen2
1. Life Science and Technology College of China Pharmaceutical University, Nanjing 210000, China;
2. Beijing Tri-Prime Gene Engineering Co Ltd, Beijing 102600, China
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Abstract  

Modified by polyethylene glycol, recombinant protein has better pharmacokinetic and pharmacodynamic properties while the structure and quality attributes become more complex. The modified recombinant protein has a big change in the structure, physical and chemical properties, and the biological activity. Therefore, it is important to consider the unique quality attributes of pegylated recombinant protein when researching the quality control. Difficulties of quality control and detection methods of important quality attributes of pegylated recombinant protein drugs are reviewed, to help future drug development and manufacturing process.



Key wordsRecombinant protein      Polyethylene glycol modification      Quality control     
Received: 06 July 2015      Published: 22 December 2015
ZTFLH:  R917  
Cite this article:

LI Xing, YAO Wen-bing, XU Chen. Quality Control for PEGylated Recombinant Protein. China Biotechnology, 2015, 35(12): 109-114.

URL:

https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20151217     OR     https://manu60.magtech.com.cn/biotech/Y2015/V35/I12/109

[1] Abuchowski A, Van Es T, Palczuk N C, et al. Alteration of immunological properties of bovine serum albumin by covalent attachment of polyethylene glycol.J Biol Chem, 1977, 252(11):3578-3581.
[2] 王旭东,李晓晖,苏志国,等.蛋白药物的聚乙二醇定点修饰策略与最佳位点.中国生物工程杂志, 2010, 30(4):101-109. Wang X D, Li X H, Su Z G, et al. Site-specific PEGylation strategies and suitable modified sites of protein drugs. China Biotechnology, 2010, 30(4):101-109.
[3] World Health Organization. WHO Guidelines on the Quality, Safety, and Efficacy of Biological Medicinal Products Prepared by Recombinant DNA Technology. Switzerland:WHO Press,2013.91-92.
[4] 梁凌宇,王婉茹,郭玉婷,等. RP-HPLC法检测CN10蛋白的PEG化修饰率及含量.微生物学免疫学进展, 2015, 43(1):14-18. Liang L Y, Wang W R, Guo Y T, et al. Determination of modification rate and content of PEGylated CN10 by RP-HPLC. Progress in Microbiology and Immunology, 2015, 43(1):14-18.
[5] 李晶,王永利,杨化新,等.三硝基苯磺酸法测定聚乙二醇化重组人生长激素的平均修饰度.现代生物医学进展, 2011,11(9):1609-1612. Li J, Wang Y L, Yang H X, et al. Determination of average degree of modification of pegylated recombinant human growth hormone by trinitrobenzene sulfonic acid. Progress in Modern Biomedicine, 2011,11(9):1609-1612.
[6] Bullock J, Chowdhury S, Severdia A, et al. Comparison of results of various methods used to determine the extent of modification of methoxy polyethylene glycol 5000-modified bovine cupri-zinc superoxidedismutase. Anal Biochem, 1997, 254(2):254-262.
[7] Stocks S J, Jones A J, Ramey C W, et al. A fluorometric assay of the degree of modification of protein primary amines with polyethylene glycol. Anal Biochem, 1986,154(1):232-234.
[8] 李晶,何辉,程速远,等.荧光胺衍生化法测定3种聚乙二醇化重组人生长激素的平均修饰率.药物分析杂志,2014,34(8):1368-1373. Li J, He H, Cheng S Y, et al. A fluorometric assay to determine the average modification rate of three kinds of PEG-rhGHs. Chin J Pharm Anal, 2014,34(8):1368-1373.
[9] 李晶,梁成罡.聚乙二醇化重组人生长激素质控难点探讨.中国新药杂志,2014,23(3):271-274. Li J, Liang C G. Difficulties in quality control of PEGylated rhGH. Chinese Journal of New Drugs,2014,23(3):271-274.
[10] 李修乐.聚乙二醇化蛋白的修饰位点识别研究.泰安:山东农业大学食品科学与工程学院, 2013. Li X L. Methods for identification of conjugationsite on pegylated protein. Taian:Institute of Food Science and Engineering, Shandong Agricultural University, 2013.
[11] Monkarsh S P, Ma Y, AglioneA,et al. Positional isomers of monopegylated interferon alpha-2a:isolation, characterization, and biological activity.Anal Biochem, 1997,247(2):434-440.
[12] Yu W L, Yu C M, Wu L, et al. PEGylated recombinant human interferon-x as a long-acting antiviral agent:Structure, antiviral activity and pharmacokinetics. Antiviral Research, 2014(108):142-147.
[13] Lu X J, Gough P C, DeFelippis M R, et al. Elucidation of PEGylation site with a combined approach of In-source fragmentation and CID MS/MS. J Am Soc Mass Spectrom, 2010, 21(5):810-818.
[14] Yoo C, Suckau D, Sauerland V, et al. Toward top-down determination of PEGylation site using MALDI in-source decay MS analysis. J Am Soc Mass Spectrom,2009,20(2):326-333.
[15] 张苏,张培彪.聚乙二醇化重组人干扰素β-1a注射液中游离聚乙二醇残留量的测定.中国现代应用药学,2014,31(8):966-968. Zhang S, Zhang P B. Determination of Free PEG in PEG-rhIFN-β-1a injection. Chin J Mod Appl Pharm, 2014,31(8):966-968.
[16] 李永红,李响,韩春梅,等.高效液相色谱蒸发光散射检测法测定聚乙二醇干扰素α-2a注射液中游离聚乙二醇的含量.药物生物技术, 2014,21(4):353-354. Li Y H, Li X, Han C M, et al. Quantitation of free polyethylene glycol in PEGylated interferon alpha-2a injection by HPLC-ELSD. Pharmaceutical Biotechnology, 2014,21(4):353-354.
[17] 苑方圆,李晶,梁成罡,等.聚乙二醇(PEG)化尿酸氧化酶中游离PEG的含量测定.药物分析杂志,2014,34(3):461-464. Yuan F Y, Li J, Liang C G, et al. Determination of the content of free polyethylene glycol in PEGylated uricase. Chin J Pharm Anal, 2014,34(3):461-464.
[18] Kurfürst M M. Detection and molecular weight determination of polyethylene glycol-modified hirudin by staining after sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Anal Biochem, 1992, 200(2):244.
[19] Hardy E, Ramón J, Saez V, et al. Detection of PEGylated proteins in polyacrylamide gels by reverse staining withzinc and imidazole salts. Electrophoresis, 2008, 29(11):2363-2371.
[20] Fee C J, Van J M. Prediction of the viscosity radius and the size exclusion chromatography behavior of PEGylatedproteins. Bioconjug Chem, 2004,15(6):1304-1313.
[21] 王军志.生物技术药物研究开发与质量控制.第2版.北京:科学出版社, 2007.71-102. Wang J Z. The Research Development and Quality Control of Biomedical. 2nd ed. Beijing:Science Press, 2007.71-102.
[22] Zheng C Y, Ma G H, Su Z G. Native PAGE eliminates the problem of PEG-SDS interaction in SDS-PAGE and provides an alternative to HPLC in characterization of protein PEGylation. Electrophoresis, 2007, 28(16):2801-2807.
[23] Möller I, Thomas A, Geyer H, et al. Synthesis, characterisation, and mass spectrometric detection of a pegylated EPO-mimetic peptide for sports drug testing purposes. Rapid Communications in Mass Spectrometry, 2011,25(15):2115-2123.
[24] 吴影新,翟艳琴,雷建都,等.聚乙二醇修饰蛋白体系的SDS-聚丙烯酰胺凝胶电泳染色方法新探.化学通报,2005,72(5):459-462. Wu Y X, Zhai Y Q, Lei J D, et al. Application of three kinds of staining methods for PEGylated protein system in SDS-PAGE. Chemistry, 2005,72(5):459-462.
[25] 谭和平,王彧婕,邹燕,等.蛋白质分子质量测试方法概述.中国测试,2011, 37(2):34-37. Tan H P, Wang Y J, Zou Y, et al. Methods of molecular weight determination of proteins. China Measurement & Test, 2011,37(2):34-37.
[26] Huang L, Gough P C, Defelippis M R. Characterization of poly(ethylene glycol)and PEGylated products by LC/MS with postcolumn addition of amines. Anal Chem, 2009, 81(2):567-577.
[27] Foser S, Schacher A, Weyer K A, et al. Isolation, structural characterization, and antiviral activity of positional isomers of monopegylated interferon α-2a(PEGASYS). Protein Expression and Purification, 2003,30(1):78-87.
[28] Huang Y S, Wen X F, Wu Y L, et al. Engineering a pharmacologically superior form of granulocytecolony-stimulating factor by fusion with gelatin-like-protein polymer. Eur J Pharm Biopharm, 2010, 74(3):435-441.
[29] Wadhwa M, Bird C, Dougall T, et al. Establishment of the first international standard for PEGylated granulocyte colony stimulating factor (PEG-G-CSF):report of an international collaborative study. Journal of Immunological Methods, 2015,416:17-28.

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