| 陈方,宋海峰,尚明美,等.125I-聚乙二醇化胸腺素α1的制备与纯化[J].中华放射医学与防护杂志,2005,25(6):505-508.CHEN Fang,SONG Hai-feng,SHANG Ming-mei,et al.Preparation and purification of 125I-labelled PEGylated thymosin α1[J].Chin J Radiol Med Prot,2005,25(6):505-508 |
| 125I-聚乙二醇化胸腺素α1的制备与纯化 |
| Preparation and purification of 125I-labelled PEGylated thymosin α1 |
| 投稿时间：2005-01-24 |
| DOI： |
| 中文关键词: 胸腺素 聚乙二醇化 125I标记 酶免疫分析方法 |
| 英文关键词:Thymosin α1 PEGylation 125I labelling Purification Enzyme immunoassay (EIA) |
| 基金项目:国家863计划重大专项(2003AA2Z347B);国家自然科学基金资助项目(39930180) |
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| 中文摘要: |
| 目的 制备高纯度放射性125I标记的酪氨酸聚乙二醇化胸腺素α1(Tyr-PEG-TA1)用于动物药代动力学研究。方法 改良Iodogen法进行125I标记。Ziptip头监测标记反应进程以优化反应条件。采用Sephadex G50与Sephadex G10两步凝胶层析法对标记混合物进行纯化。利用酶免疫分析(EIA)方法鉴定PEG-TA1增加酪氨酸残基前后以及Tyr-PEG-TA1经125I标记前后免疫学活性的变化。结果 PEG-TA1增加酪氨酸残基前后以及Tyr-PEG-TA1经125I标记前后免疫活性均无明显变化,纯化后的标记产物放化纯度为95.9%,放射性比活度为39.4Bq/ng。结论 成功地制备了放化纯度和放射性比活度均满足药代动力学研究的125I-Tyr-PEG-TA1。 |
| 英文摘要: |
| Objective To prepare a highly purified 125I-pegylated thymosin α1 (PEG-TA1) with satisfied bioactivity for using in the distribution and excretion study in animals. Methods Tyr-PEG-TA1 was iodinated by means of Iodogen method. The ZipTip pipette tips were utilized to monitor the reaction process for the optimization of reaction conditions. A two-step gel filtration chromatography with Sephadex G50 followed by Sephadex G10 was applied to purify the labelled products. The bioactivity assay of PEG-TA1, Tyr-PEG-TA1 and 125I-Tyr-PEG-TA1 were performed by an enzyme immunoassay (EIA) method. Results The optimum condition of labelling reaction was: shaking for 20 min at 25℃ in 0.1 mol/L phosphate buffer (0.1mol/L NaCl) with the pH value of 7.0. Slight but no significant difference of bioactivity among the PEG-TA1, Tyr-PEG-TA1 and the 125I-Tyr-PEG-TA1 was observed. After the two-step purification process, a product of 125I-Tyr-PEG-TA1 with radiochemical purity of 95% and specific activity of 39.4 Bq/ng was obtained. Conclusion A successful preparation of 125I-Tyr-PEG-TA1 was achieved, and the radiochemical purity and the specific activity of the 125I labeled molecules satisfied the requirements of non-clinical pharmacokinetic study. |
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