An improved synthesis of NHS-MAG3 for conjugation and radiolabeling of biomolecules with 99mTc at room temperature
An improved synthesis of NHS-MAG3 for conjugation and radiolabeling of biomolecules with 99mTc at room temperature
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DOI：
10.1038/nprot.2007.144
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发表时间：
2007-01-01
期刊：
影响因子：
14.8
通讯作者：
Hnatowich, Donald J.
中科院分区：
文献类型：
--
作者：
Wang, Yi;Liu, Xinrong;Hnatowich, Donald J.
巯基乙酰基三甘氨酸（MAG 3）螯合剂已被证明可以稳定地络合用于核成像的锝-99m（Tc-99 m）和用于肿瘤放射治疗研究的放射性铼（Re-186/188）。具有S-乙酰基保护的MAG 3的双官能N-羟基琥珀酰亚胺酯（N-羟基琥珀酰亚胺基S-乙酰基巯基乙酰基三甘氨酸酯（NHS-MAG 3））已成功地用于将MAG 3螯合剂共价缀合至伯胺官能化的生物分子。我们在此描述了NHS-MAG 3的简化合成，其开始于由巯基乙酸制备S-乙酰巯基乙酸的N-羟基琥珀酰亚胺酯（N-琥珀酰亚胺基S-乙酰巯基乙酸酯（SATA）），然后由SATA合成S-乙酰巯基乙酰三甘氨酸，总共需要约14天。最后，从S-乙酰巯基乙酰基三甘氨酸合成NHS-MAG 3需要另外5天。我们先前描述了一种用于制备MAG 3缀合和Tc-99 m放射性标记的生物分子的方法，该方法在缀合后纯化期间需要升高的温度。我们现在报告一种改进的制备方法，该方法在室温下完成，因此适用于温度敏感的生物分子。牛血清白蛋白的偶联和放射性标记用作示例。偶联和纯化需要约2-3小时，放射性标记和标记后纯化需要约另外2小时。
The mercaptoacetyltriglycine (MAG3) chelator has been shown to stably complex technetium-99m (Tc-99m) for nuclear imaging and radiorhenium (Re-186/188) for tumor radiation therapy studies. The bifunctional N-hydroxysuccinimidyl ester of MAG3 with S-acetyl protection (N-hydroxysuccinimidyl S-acetylmercaptoacetyltriglycinate (NHS-MAG3)) has been successfully used to covalently conjugate a MAG3 chelator to primary amine functionalized biomolecules. We describe herein a simplified synthesis of NHS-MAG3 that begins with the preparation of the N-hydroxysuccinimidyl ester of S-acetylmercaptoacetic acid (N-succinimidyl S-acetylmercaptoacetate ( SATA)) from mercaptoacetic acid and is followed by the synthesis of S-acetylmercaptoacetyltriglycine from SATA, together requiring about 14 days. Finally, the synthesis of NHS-MAG3 from S-acetylmercaptoacetyltriglycine requires a further 5 days. We had earlier described a method for the preparation of MAG3-conjugated and Tc-99m-radiolabeled biomolecules that required elevated temperatures during postconjugation purification. We now report a modified method for the preparation that is accomplished at room temperature and therefore applicable to temperature-sensitive biomolecules. The conjugation and radiolabeling of bovine serum albumin is used as an example. The conjugation and purification requires about 2-3 h and the radiolabeling and postlabeling purification requires about an additional 2 h.