Pulsed ultrafiltration mass spectrometry: A new method for screening combinatorial libraries
Pulsed ultrafiltration mass spectrometry: A new method for screening combinatorial libraries
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DOI：
10.1021/ac970132j
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发表时间：
1997-06-01
影响因子：
7.4
通讯作者：
Venton, DL
中科院分区：
文献类型：
--
作者：
vanBreemen, RB;Huang, CR;Venton, DL
为了满足药物发现过程中快速筛选组合文库以鉴定新的先导化合物的需求，我们开发了超滤和电喷雾质谱的在线组合，称为脉冲超滤质谱，它有助于识别文库混合物中与溶液相受体结合的溶液相配体。将文库混合物中的配体与大分子受体（例如人血清白蛋白或小牛肠腺苷脱氨酶）结合后，通过超滤纯化配体-受体复合物，然后使用甲醇解离，将配体洗脱到电喷雾质谱仪中进行检测。利用该方法成功地结合、释放和检测了解离常数在微摩尔至纳摩尔范围内的配体，包括与人血清白蛋白结合的华法林、水杨酸盐、呋塞米和甲状腺素，以及与小牛肠腺苷脱氨酶结合的erythro-9-(2-羟基-3-壬基)腺嘌呤。重复的结合和释放实验表明受体可以重复使用，因此，脉冲超滤质谱法被证明为筛选组合文库提供了一种简单而强大的新方法，以支持新药的发现。
In response to the need for rapid screening of combinatorial libraries to identify new lead compounds during drug discovery, we have developed an on-line combination of ultrafiltration and electrospray mass spectrometry, called pulsed ultrafiltration mass spectrometry, which facilitates the identification of solution-phase ligands in library mixtures that bind to solution-phase receptors. After ligands contained in a library mixture were bound to a macromolecular receptor, e.g., human serum albumin or calf intestine adenosine deaminase, the ligand-receptor complexes were purified by ultrafiltration and then dissociated using methanol to elute the ligands into the electrospray mass spectrometer for detection. Ligands with dissociation constants in the micromolar to nanomolar range were successfully bound, released, and detected using this method, including warfarin, salicylate, furosemide, and thyroxine binding to human serum albumin, and erythro-9-(2-hydroxy-3-nonyl)adenine binding to calf intestine adenosine deaminase. Repetitive bind-and-release experiments demonstrated that the receptor could be reused, Thus, pulsed ultrafiltration mass spectrometry was shown to provide a simple and powerful new method for the screening of combinatorial libraries in support of new drug discovery.