A robust small-molecule microarray platform for screening cell lysates
A robust small-molecule microarray platform for screening cell lysates
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DOI：
10.1016/j.chembiol.2006.03.004
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发表时间：
2006-05-01
影响因子：
--
通讯作者：
Koehler, Angela N.
中科院分区：
文献类型：
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作者：
Bradner, James E.;McPherson, Olivia M.;Koehler, Angela N.
在这里，我们报告的小分子微阵列，包括固定的伯醇，仲醇，酚，羧酸，异羟肟酸，硫醇和胺在一个单一的载玻片表面上的扩展的官能团的兼容性。通过使用异氰酸酯介导的共价捕获策略，产生了包含近10，000种已知生物活性小分子、天然产物和源自多种多样性定向合成的小分子的小分子“多样性微阵列”。用针对感兴趣的化合物的抗体检测选择的打印的生物活性化合物。多样性微阵列的新表面与涉及细胞裂解物的方法高度兼容。这一特征使得使用细胞裂解物的稳健的优化筛选方法成为可能，允许通过使用表位标记的或嵌合的荧光蛋白而无需预先纯化来检测具有广泛结合亲和力的特异性相互作用。我们相信，这种扩大的研究能力在生物学和医学方面具有相当大的前景。
Herein we report the expanded functional group compatibility of small-molecule microarrays to include immobilization of primary alcohols, secondary alcohols, phenols, carboxylic acids, hydroxamic acids, thiols, and amines on a single slide surface. Small-molecule "diversity microarrays" containing nearly 10,000 known bioactive small molecules, natural products, and small molecules originating from several diversity-oriented syntheses were produced by using an isocyanate-mediated covalent capture strategy. Selected printed bioactive compounds were detected with antibodies against compounds of interest. The new surface of the diversity microarrays is highly compatible with approaches involving cellular lysates. This feature has enabled a robust, optimized screening methodology using cellular lysates, allowing the detection of specific interactions with a broad range of binding affinity by using epitope-tagged or chimeric fluorescent proteins without prior purification. We believe that this expanded research capability has considerable promise in biology and medicine.