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Synthesis and screening of a combinatorial peptide library for ligands to target transferrin: Miniaturizing the library.

机译：合成和筛选针对目标转铁蛋白的配体的组合肽文库：将文库微型化。

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Combinatorial libraries are used in the search for ligands that bind to target proteins. Fmoc solid-phase peptide synthesis is routinely used to generate such libraries. Microwave-assisted peptide synthesis was employed here to decrease reaction times by 80-90%. Two One-Bead-One-Compound combinatorial libraries were synthesized on 130mum beads (one containing 750 members and the other 16, 807). The use of smaller solid supports would have many important practical advantages including; increased library diversity per unit mass, smaller quantities of library needed to generate hits, and screening could be conducted by using a standard flow cytometer. To this end, a miniaturized peptide library was synthesized on 20 mum beads to demonstrate proof of principle.;A small sample from the 16,807-member library was screened against transferrin-AlexaFluro 647, a protein responsible for iron transport in vivo. A number of hits were identified and sequenced using techniques coupling nanomanipulation with nanoelectrospray mass spectrometry.

机译：组合文库用于寻找与靶蛋白结合的配体。 Fmoc固相肽合成通常用于生成此类文库。此处采用微波辅助肽合成将反应时间减少80-90％。在130mum的微珠上合成了两个单珠一化合物组合文库（一个包含750个成员，另一个包含16个，807）。使用较小的固体载体将具有许多重要的实际优势，包括；每单位质量的文库多样性增加，产生命中所需的文库数量减少，可以使用标准流式细胞仪进行筛选。为此，在20个妈妈的珠子上合成了一个微型化的肽库，以证明其原理性。筛选了来自16,807个成员库的一小部分样本，以检测转铁蛋白-AlexaFluro 647（一种负责体内铁转运的蛋白质）。使用结合了纳米处理和纳米电喷雾质谱的技术，鉴定了许多命中并进行了测序。

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作者
Brown, Jennifer M.;
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University of North Texas.;

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授予单位 University of North Texas.;
学科 Chemistry Biochemistry.;Chemistry Organic.
学位 M.S.
年度 2010
页码 71 p.
总页数 71
原文格式 PDF
正文语种 eng
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Synthesis and screening of a combinatorial peptide library for ligands to target transferrin: Miniaturizing the library.

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