Wu Yan-Qi, Yang Fei-Peng, Wang Hong-Yin, Liu Jian-Xin, Liu Zheng-Chun
School of Geosciences and Info-Physics, Institute of Biomedical Engineering, Central South University, Changsha 410083, China.
J Nanosci Nanotechnol. 2013 Mar;13(3):2061-7. doi: 10.1166/jnn.2013.6890.
Peptide nucleic acids (PNA) microarray assembled with hundreds of unique PNA oligomers has been regarded as a new and mighty competitor of DNA chip in gene analyzing. However, PNA microarray is still a luxury art due to the difficult and laborious chemical synthesis. Herein, we have developed a fully-automated synthesizer for PNA microarray through photolithography. A preactivation mixer was designed and integrated into the synthesizer in order to get rid of the annoying manual process and increase the coupling efficiency of PNA monomers. The PNA patterning model was carried out to check the performance of the automated synthesizer, revealing that an exposure time of 3 min was sufficient for the complete removal of o-nitroveratryloxycarbonyl (NVOC) groups from the synthetic sites with the help of photosensitizer isopropylthioxanthone and the stepwise yield was measured to be about 98.0%, which is comparable with that from conventional fluorenyl-methyloxycarbonyl (FMOC) chemistry. Those results have definitely demonstrated the possibility and capability of this fully-automated synthesizer to fabricate high-quality PNA microarrays.
由数百种独特的肽核酸(PNA)寡聚物组装而成的PNA微阵列,在基因分析中被视为DNA芯片的一种强大的新竞争对手。然而,由于化学合成困难且费力,PNA微阵列仍然是一种昂贵的技术。在此,我们通过光刻技术开发了一种用于PNA微阵列的全自动合成仪。设计并将一个预活化混合器集成到合成仪中,以消除繁琐的手动操作过程并提高PNA单体的偶联效率。进行了PNA图案化模型以检查自动合成仪的性能,结果表明,在光敏剂异丙基硫代呫吨酮的帮助下,3分钟的曝光时间足以完全去除合成位点上的邻硝基藜芦氧基羰基(NVOC)基团,逐步产率测得约为98.0%,这与传统的芴甲氧羰基(FMOC)化学方法相当。这些结果明确证明了这种全自动合成仪制造高质量PNA微阵列的可能性和能力。