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在蛋白质及多肽等生物大分子研究中,往往需要用放射性同位素进行标记以达到追踪或检测的目的。1981年Dolly等报道用N-琥珀酰亚胺[2,3-~3H]丙酸-酯(~3H-NSP)标记银环蛇毒素成功,1983年Yuis等发表了合成~3H-NSP的方法。我们参考此法,改进了反应溶剂和纯化方法,合成了~3H-NSP,总产率17.3％,比度525.4GBq/mmol,放化纯>98％,随后又将丙烯酸与N-羟基琥珀酰亚胺先缩合成琥珀酰亚胺丙烯酸酯,再氚化制备~3N-NSP,获得成功,产率80％,比度1.33TBq/mmol,放化纯>95％,从而使引入标记元素改在最后一步,缩小了合成量,节省了氚气,并提高了产率。用我们合成的~3H-NSP分别标记了丙种球蛋白、乙酰胆碱酯酶、过氧化氢酶和眼镜蛇毒素都获成功,并保持生物活性。
In the study of biological macromolecules such as proteins and peptides, it is often necessary to label them with radioisotopes for tracking or detection purposes. In 1981, Dolly et al. Reported the successful labeling of silver bungarotoxin with N-succinimidyl [2,3- ~ 3H] propionate (~ 3H-NSP). In 1983, Yuis et al. Published a method for the synthesis of ~ 3H-NSP . With reference to this method, the reaction solvent and the purification method were improved, ~ 3H-NSP was synthesized with a total yield of 17.3%, a specific gravity of 525.4 GBq / mmol and a radiochemical purity of> 98%. Subsequently, acrylic acid and N-hydroxysuccinimide The imine was first condensed to succinimidyl acrylate and then tritiated to produce ~ 3N-NSP with a yield of 80%, a specific gravity of 1.33 TBq / mmol and a radiochemical purity of> 95% The last step, reducing the amount of synthesis, saving tritium gas, and increase the yield. Hg-globulin was labeled with ~ 3H-NSP synthesized by us. Both acetylcholinesterase, catalase and cobra venom were successful and maintained their bioactivity.