Experimental study of triplex-forming oligonucleotide targeted to the initiator of S gene of HBV labeled with ¹²⁵I

Experimental study of triplex-forming oligonucleotide targeted to the initiator of S gene of HBV labeled with 125 I 1 first-author 1 2 1 1 1 Shanghai 10th People's Hospital, Tongji University, Shanghai 200072, China; Shanghai 10th People's Hospital, Tongji University, Shanghai 200072, China; School of Life Science & Technology, Tongji University, Shanghai 200092, China School of Life Science & Technology, Tongji University, Shanghai 200092, China This study is used to investigate the feasibility of employing the Iodogen method to label triplex-forming oligonucleotide (TFO) targeted to the initiator of the S gene of HBV with 125 I. A 17-mer oligonucleotides sequence was synthesized and grafted at the 5＇ terminal with a tyramine group. Radioiodination of the tyramine-TFO with 125 I was then performed using the Iodogen method. After TFO was labeled with 125 I using the Iodogen method, the labeling rate, the radiochemical purity, stability and bioactivity were determined, respectively. The results show that the radiolabeling rate and the radiochemical purity were 93% and 99%, respectively; and the radiochemical purity is more than 90% in vitro at -20℃ on the 5th day after labeling; and the rate of 125 I-tyramine-TFO binding to HepG2.2.15 cells was (37.2±1.4) % and statistically different from the rate of HepG2 ( p ﹤0.5). Hence, it is concluded that the labeling of oligonucleotides conjugated with tyramine using the Iodogen method is successful and is characterized with a high labeling rate, high stability, and a low loss of bioactivity of the labeled agent. Triplex-forming oligonucleotide(TFO) 125 I Labeling References Panyutin IG , Winters T A , Feinendegen L E , et al . The Quarterly Journal of Nuclear Medicine , 2000 , 44 : 256 - 267 The Quarterly Journal of Nuclear Medicine Martin R F , Haseltine W A . Science , 1981 , 213 ( 4510 ): 896 - 898 . Science England P R , Murray V . Antisense Res ., 1993 , 3 ( 2 ): 219 - 224 . Antisense Res Panyutin I G , Neumann R D . Nucleic Acids Res ., 1994 , 22 ( 23 ): 4979 - 4982 . Nucleic Acids Res Lu Changde , Chen Jinhui , Chuang Min , et al . Chinese Patent No. 97 1 06667.1 . Panyutin I , Neumann R . Acta Oncol , 1996 , 35 : 817 - 823 . Acta Oncol Commerford S L . Biochem , 1971 , 10 : 1993 - 1999 . Biochem Dewanjee M K , Ghafouripour A K , Werner R K , et al . Bioconjug Chem , 1991 , 2 ( 4 ): 195 - 200 . Bioconjug Chem Liu Bingchen , Yue Jinyin , Mo Chuangjie , et al . Chinese Journal of Nuclear Medicine . 2000 , 20 ( 1 ): 33 - 34 . Chinese Journal of Nuclear Medicine Cammilleri S , Sangrajrang S , Perdereau B , et al . Eur J Nucl Med , 1996 , 23 ( 4 ): 448 - 452 . Eur J Nucl Med Arimondo P B , Barcelo F , Sun J S , et al . Biochemistry . 1998 , 37 ( 47 ): 16627 - 16635 . Biochemistry Hanotowich D J . J Nucl Med , 1999 , 40 : 693 - 703 . J Nucl Med 10.1016/S1001-8042(06)60030-4 R817 Supported by National Natural Science Foundation of China (No.30300092) . 2006-01-26 2006-06-20 2021-01-19 2006 Nuclear Science and Techniques 03 17