Combinatorial Cellulose-Bound Peptide Libraries: Screening Tools for the Identification of Peptides That Bind Ligands with Predefined Specificity
Combinatorial Cellulose-Bound Peptide Libraries: Screening Tools for the Identification of Peptides That Bind Ligands with Predefined Specificity
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组合纤维素结合肽文库：用于鉴定以预定特异性结合配体的肽的筛选工具
DOI：
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发表时间：
1994
期刊：
影响因子：
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通讯作者：
J. Schneider
中科院分区：
文献类型：
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作者：
A. Kramer;A. Schuster;U. Reineke;R. Malin;R. Volkmer‐Engert;C. Landgraf;J. Schneider
摘要我们描述了在连续纤维素膜支持物上合成结构不同类型的组合肽库。这些由数千万种不同肽组成的文库被筛选其结合给定配体的能力，所述配体例如单克隆抗体Tab 2、转化生长因子-β（TGFβ）、镍（II）（Ni 2+）和锝-99m（99 m Tc）。因此，我们能够检测Tab 2识别的线性转化生长因子-α（TFGα）表位SHFND。在同一实验中还鉴定了其他也结合Tab 2的肽。用于鉴定与生物学感兴趣的其它配体结合的肽混合物的第一筛选步骤显示为与Ni 2+和99 m Tc结合的肽混合物XB 1 XB 2 XX（B =确定的氨基酸，X =随机化位置）。组合线性全L-或全D-文库XXB 1 B 2 XX和两个文库通过C-和N-末端半胱氨酸之间的二硫键[环状]构象上受到限制，（C1-C8）-C1 XXB 1 B 2 XXC 8 ]或N-末端的α氨基和C-末端谷氨酸的γ羧基之间的酰胺键[环（X 1 -E 7）-X 1 X B 1 B 2 XXE 7 ]用转化生长因子-β筛选，得到与该配体结合的结构不同的肽混合物。所获得的结果表明，化学上不同类型的纤维素结合的组合文库可以很容易地制备，允许快速和廉价的筛选数百万肽的选择与给定的配体，如蛋白质，金属，核酸，和其他生物学感兴趣的分子结合的预定义的特异性的单个分子。
Abstract We describe the synthesis of structurally different types of combinatorial peptide libraries on continuous cellulose membrane supports. These libraries consisting of tens of millions of different peptides were screened for their ability to bind given ligands such as the monoclonal antibody Tab2, transforming growth factor-β (TGFβ), nickel(II) (Ni 2+ ), and technetium-99m ( 99m Tc). We were thus able to detect the linear transforming growth factor-α (TFGα) epitope SHFND recognized by Tab2. Other peptides that also bound Tab2 were identified within the same experiment. A first screening step for identification of peptide mixtures that bind to other ligands of biological interest is shown for peptide mixtures XB 1 XB 2 XX (B = defined amino acid, X = randomized position) that bind to Ni 2+ and 99m Tc. A combinatorial linear all L- or all D-library XXB 1 B 2 XX and two libraries conformationally restrained either via a disulfide bridge between a C-and an N-terminal cysteine [cyclo(C 1 -C 8 )-C 1 XXB 1 B 2 XXC 8 ] or an amide bond between the alpha amino group of the N-terminus and the gamma carboxyl group of a C-terminal glutamic acid [cyclo(X 1 -E 7 )-X 1 XB 1 B 2 XXE 7 ] were screened with transforming growth factor-β, resulting in structurally different peptides mixtures that bound to this ligand. The results obtained indicate that chemically different types of cellulose-bound combinatorial libraries can be prepared easily, allowing the rapid and inexpensive screening of millions of peptides for selection of single molecules with predefined specificity that bind to given ligands such as proteins, metals, nucleic acids, and other molecules of biological interest.