聚乙二醇化重组人生长激素的性质及活性 |
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引用本文: | 魏练平 王荣海 戎隆富 宋礼华. 聚乙二醇化重组人生长激素的性质及活性[J]. 中国生物制品学杂志, 2005, 18(2): 138-140 |
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作者姓名: | 魏练平 王荣海 戎隆富 宋礼华 |
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作者单位: | [1]安徽大学生命科学学院,合肥230039 [2]安徽省生物研究所,合肥230088 |
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基金项目: | 2 0 0 4~ 2 0 0 5年度安徽省优秀青年科技基金项目 (编号0 40 43 0 75 )资助 . |
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摘 要: | 目的 研究聚乙二醇化重组人生长激素的性质及活性。方法 通过分子筛高效液相 (SizeexclusionHPLC)、毛细管等电聚焦 (CIEF)、圆二色谱 (CD)以及荧光光谱等方法检测单聚PEG- GH的纯度、等电点、二级结构以及分子构象 ;采用妊娠兔肝细胞放射受体分析法 (RRA)检测单聚PEG GH的体外受体亲和力 ,采用未成年去垂体大鼠体重增加法检测体内生物活性。结果 分子筛高效液相图谱上仅出现一个峰 ,表明样品纯度很高 ,等电点范围为 5 .1~ 5 .2 ,与未修饰的rhGH的等电点相一致 ;CD谱与荧光光谱均表明单个mPEG ALD的修饰对rhGH的二级结构以及分子构象没有明显影响 ;与未修饰的rhGH相比 ,单聚PEG GH体外受体亲和力明显降低 ;但体内作用时间更长 ,且更加有效。结论 此结果为进一步研究和开发长效重组人生长激素提供了理论依据。
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关 键 词: | 重组人生长激素 聚乙二醇丙醛 化学修饰 放射受体分析法 受体亲和力 生物活性 |
修稿时间: | 2004-11-15 |
Characteristics and Biological Activity of Mono-PEG-GH |
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Abstract: | Objective To study the charactristics and bio logi cal activity of PEG-modified recombinant human growth hormone(mono-PEG-GH). Methods The purity,isoelectric point,secondary structure and mole cular conformation of mono-PEG-GH were analyzed by the methods including size exclusion HPLC,capillary isoelectric focusing,circular dichrosim(CD) and fluores cence spectrometry.The receptor binding activity in vitro of mono-PEG-GH w as e valuated by analyzing its inhibitory effect on the binding of 125I-hG H to growt h hormone receptor on the membrane of pregnant rabbit liver cells using radiorec eptor assay.The biological activity in vivo of mono-PEG-GH was analyzed by bod y weight gain test in hypophysectomized rats.Results High purit y of mono-PEG-GH was proved by the only peak on size exclusion HPLC profile.The isoelectric point of mono-PEG-GH was 5 1-5 2,which was consistent with tha t of unmodified rhGH.Neither CD nor fluorescence spectrometry showed the influen ce of modification with mPEG-ALD on the secondary structure or molecular confor mation of rhGH.Compared with that of unmodified rhGH,the receptor affinity in vi tro of mono-PEG-GH decreased significantly.However,its receptor affinity i n vivo was more persistent and effective.Conclusion It provid ed a theoretical basis for the study and development of long-acting rhGH. [ |
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Keywords: | Recombinant human growth hormone mPEG-ALD Chemical modification Receptor aff inity Biological activity |
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