Antibody-nucleotide conjugate as a substrate for DNA polymerases.
Antibody-nucleotide conjugate as a substrate for DNA polymerases.
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DOI：
10.1039/c8sc01839a
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发表时间：
2018-09-21
期刊：
影响因子：
8.4
通讯作者：
Marx A
中科院分区：
文献类型：
--
作者：
Balintová J;Welter M;Marx A
抗体修饰的核苷酸被开发出来，并通过DNA聚合酶将它们掺入新生的DNA中，用于发展DNA的肉眼检测。在这里，我们报道了一种抗体修饰的核苷酸的发展及其在DNA聚合酶催化下选择性地掺入新生的DNA中。虽然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，但它被酶很好地接受。此外，抗体的识别不会因结合而取消，但可以被与信号产生酶(即马萝卜过氧化物酶)结合的二次抗体识别。因此，这种产品可以用肉眼对核苷酸掺入进行比色读出，从而可以检测到低至10amol的DNA。在未来，像这里描述的那样的分析可能允许在没有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。
Antibody-modified nucleotides are developed and their incorporation into nascent DNA by DNA polymerases exploited for the development of the naked-eye detection of DNA. 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.
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影响因子：
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