| CHEN Fang,SONG Hai-feng,SHANG Ming-mei.Preparation and purification of 125I-labelled PEGylated thymosin α1[J].Chinese Journal of Radiological Medicine and Protection,2005,25(6):505-508 |
| Preparation and purification of 125I-labelled PEGylated thymosin α1 |
| Received:January 24, 2005 |
| DOI： |
| KeyWords:Thymosin α1 PEGylation 125I labelling Purification Enzyme immunoassay (EIA) |
| FundProject:国家863计划重大专项(2003AA2Z347B);国家自然科学基金资助项目(39930180) |
| Author Name | Affiliation | | CHEN Fang | Institute of Radiation Medicine, Academy of Military Medical Sciences, Beijing 100850, China | | SONG Hai-feng | Institute of Radiation Medicine, Academy of Military Medical Sciences, Beijing 100850, China | | SHANG Ming-mei | Institute of Radiation Medicine, Academy of Military Medical Sciences, Beijing 100850, China | | 欧伦 | Institute of Radiation Medicine, Academy of Military Medical Sciences, Beijing 100850, China | | 朱宝珍 | Institute of Radiation Medicine, Academy of Military Medical Sciences, Beijing 100850, China | | 孙效 | Institute of Radiation Medicine, Academy of Military Medical Sciences, Beijing 100850, China | | 刘秀文 | Institute of Radiation Medicine, Academy of Military Medical Sciences, Beijing 100850, China |
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| Abstract:: |
| 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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