High-Throughput Measurement of Binding Kinetics by mRNA Display and Next-Generation Sequencing.
High-Throughput Measurement of Binding Kinetics by mRNA Display and Next-Generation Sequencing.
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DOI：
10.1002/anie.201600077
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发表时间：
2016-03-14
期刊：
影响因子：
--
通讯作者：
Roberts RW
中科院分区：
文献类型：
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作者：
Jalali-Yazdi F;Lai LH;Takahashi TT;Roberts RW
对表征配体亲和力的高通量方法有很大的需求。通过将 mRNA 展示与下一代测序相结合，我们确定了超过两万个配体的动力学结合和解离速率，而无需合成或纯化单个成员。我们的结果具有可重复性，并且与其他亲和力测量方法一样准确。我们能够获得超过两万个单独配体的靶蛋白的动力学结合和解离速率，而无需单独合成每个单独的配体。我们通过结合 mRNA 展示和高通量 DNA 测序来实现这一目标
There is great demand for high-throughput methods to characterize ligand affinity. By combining mRNA display with next-generation sequencing, we determined the kinetic on- and off-rates for over twenty thousand ligands, without the need for synthesis or purification of individual members. Our results are reproducible and as accurate as other methods of affinity measurement. We were able to obtain the kinetic on- and off-rates of over twenty thousand individual ligands for their target protein, without the need to synthesize each individual ligand separately. We accomplished this by combining mRNA display and high throughput DNA sequencing