A modified method for synthesis of [γ-32P] labelled adenosine triphosphate

Rahman, Wira Y.; Awaludin, Rohadi; Sarmini, Endang; Herlina, .; Triyanto,; Ritawidya, Rien; Mutalib, Abdul; Nurbaiti, Santi

doi:10.1007/s10967-014-3835-1

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“…Akan tetapi sampai saat ini ketersediaan senyawa tersebut masih sangat kurang. Oleh karena itu PTRR (Pusat Teknologi Radioisotop dan Radiofarmaka) -BATAN sebagai institusi pemerintah dalam bidang litbang radioisotop dan radiofarmaka mengembangkan penelitian senyawa nukleotida bertanda [α-32 P]ATP dari senyawa nukleotida bertanda *γ-32 P]ATP yang sudah berhasil dibuat oleh PTRR [6][7][8][9] .…”

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Sintesis Dan Karakterisasi Nukleotida Bertanda [Î±-32p]atp

et al. 2019

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The utilization of nuclear technology in health sector with molecular techniques is increasingly developed today, especially in Indonesia. One of which is nucleotide compound marked with [α-32P]ATP, this compound has been used as tracer for deoxyribonucleic acid (DNA)/ ribonucleic acid (RNA) in the study of various physiological and pathological processes. [α-32P]ATP is synthesized through several stages of continuous reaction in one reaction vessel. It begins with synthesis of [γ-32P]ATP through an enzymatic reaction, using H332PO4 and ADP, and enzymes of lactate dehydrogenase, 3-phosphoglycerate phosphokinase and glyceraldehide 3-phospho dehydrogenase; followed by phosphorilation of 3’AMP with T4 polinucleotide kinase enzyme to produce 3’-[5’-32P]ATP. The result is hydrolyzed with nuclease P1 enzyme to produce [5’-32P]AMP. The unreacted [γ-32P] is degraded by the addition of hexokinase enzyme and glucose. At the final stage of the reaction, the [5’-32P]AMP is phosphorilated using phosphoenol-piruvat, piruvat kinase, and myokinase to produce [α-32P]ATP. The test results show that the every stage of reaction is characterized using TLC method, PEI cellulose paper as stationary phase and KH3PO4 0,5 M pH 3,5 as mobile phase. At the end of reaction, the yield of [α-32P]ATP reaches 71,7%, at Rf = 0,2.

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“…Pada penelitian ini akan dilakukan pembuatan senyawa nukleotida bertanda [α-32 P]ATP melalui proses enzimatis senyawa nukleotida bertanda [γ-32 P]-ATP [8,9] . Reaksi enzimatis tersebut dapat dilihat pada reaksi berikut ini [10,11] :…”

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