Antibody–nucleotide conjugate as a substrate for DNA polymerases†,Chemical Science

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Antibody–nucleotide conjugate as a substrate for DNA polymerases†
Chemical Science ( IF 7.6 ) Pub Date : 2018-07-24 00:00:00 , DOI: 10.1039/c8sc01839a
J Balintová ₁ , M Welter ₁ , A Marx ₁

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Here we report on the development of an antibody-modified nucleotide and its sequence-selective incorporation into nascent DNA catalysed by DNA polymerases. Although the modification of the nucleotide is several orders of magnitude larger than the natural dNTP substrate and even exceeds the size of the DNA polymerase, it is well accepted by the enzyme. Moreover, the recognition of the antibody is not abolished by the conjugation but can be recognized by a secondary antibody that is conjugated to a signal-generating enzyme (i.e., horse radish peroxidase). This product can thus be exploited for a colorimetric read-out of nucleotide incorporation by the naked eye that allows detection of DNA as low as 10 amol. In future, assays like the one described herein might allow nucleic acid diagnostics at single nucleotide resolution without any laboratory equipment.

中文翻译：

抗体-核苷酸缀合物作为 DNA 聚合酶的底物†

在此，我们报告了抗体修饰核苷酸的开发及其在 DNA 聚合酶催化下序列选择性掺入新生 DNA 的情况。尽管核苷酸的修饰比天然 dNTP 底物大几个数量级，甚至超过 DNA 聚合酶的大小，但它很好地被酶所接受。此外，抗体的识别不会因缀合而消除，而是可以被与信号产生酶(即辣根过氧化物酶)缀合的二抗所识别。因此，该产品可用于通过肉眼比色读出核苷酸掺入情况，从而检测低至 10 amol 的 DNA。将来，像本文描述的那样的测定可能允许在不使用任何实验室设备的情况下以单核苷酸分辨率进行核酸诊断。

更新日期：2018-07-24

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