結合奈米材料偵測寡去氧核糖核苷酸和藥物複合體之質譜法

本論文是利用碲化汞奈米結構 (HgTe nanostructures) 作為基質輔助雷射脫附游離質譜法 (surface-assisted laser desorption/ionization mass spectrometry, SALDI-MS) 之基質，並應用於寡去氧核糖核苷酸 (oligodeoxynucleotides, ODNs) 及其和藥物複合體 (ODN-drug complexes) 之偵測。本研究所使用之寡去氧核糖核苷酸為四條易於急性骨髓白血病 (acute myeloid leukemia, AML) 病患中發現的序列，而藥物則是常用於治療急性骨髓白血病患的雙羥蒽醌 (Mitoxantrone, MTX)。本研究結果顯示此方法能成功地同時偵測到四條寡去氧核糖核苷酸，其質荷比 (m/z) 值分別為4571、 4586、 4610 和 4635，以及其與藥物之複合體，其質荷比值分別為5017、 5031、 5055和 5079。質荷比值在4610的寡去氧核糖核苷酸為正常表現的基因序列，而其它三條則為在急性骨髓白血病患中發生單點突變的基因序列。這方法對於此四條寡去氧核糖核苷酸所得到的偵測極限可以低至2 nM，說明其有潛力應用於急性骨髓白血病的診斷。經由此基質輔助雷射脫附游離質譜法也能計算四條寡去氧核糖核苷酸與雙羥蒽醌複合體的解離常數 (dissociation constants, Kd)，此基質輔助雷射脫附游離質譜法方法與傳統光學的吸收方法所得到的Kd值在Student’s t test的統計法上無明顯差異。相對於傳統光學方法，此SALDI-MS方法提供了簡單、快速、良好的再現性及樣品需求量少等優勢。

關鍵字

寡去氧核糖核苷酸；奈米材料；解離常數；基質輔助雷射脫附游離質譜儀

並列摘要

A surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS) using HgTe nanostructures as the matrix has been employed for the detection of four 15-base oligodeoxynucleotides (ODNs) and their complexes with mitoxantrone (MTX) that is a common drug for treatment of acute myeloid leukemia (AML) patients. The major peaks for the four tested ODNs are genes found in AML patients at m/z values of 4571, 4586, 4610, and 4635, while they are at m/z values of 5017, 5031, 5055, and 5079 for their corresponding complexes with MTX. The ODN with m/z value of 4610 is assigned for a normal gene of AML, while the other three are single-base mutant ODNs. This approach allows detection of the tested ODNs at the concentrations down to 2 nM, showing their potential for diagnosis of AML. The dissociation constants values of the four tested ODN-MTX complexes determined by the SALDI-MS approach are similar and all in the μM level, which agree with that determined by applying a conventional absorption approach. Relative to the conventional approach, the SALDI-MS approach has advantages of simplicity, rapidity, reproducibility, and use of smaller amounts of ODNs and MTX.

並列關鍵字

oligodeoxynucleotides ； nanomaterial ； dissociation constants ； surface-assisted laser desorption/ionization mass spectrometry

參考文獻

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結合奈米材料偵測寡去氧核糖核苷酸和藥物複合體之質譜法

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