Generating DNA synbodies from previously discovered peptides.
Generating DNA synbodies from previously discovered peptides.
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DOI：
10.1002/cbic.201100284
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发表时间：
2011-08-16
期刊：
影响因子：
3.2
通讯作者：
Chaput, John C.
中科院分区：
文献类型：
--
作者：
Liu, Rui;Jiang, Bing;Yu, Hanyang;Chaput, John C.
在蛋白质组范围内创建蛋白质捕获试剂是分子医学中的一个巨大挑战。[1]高质量的试剂对于阐明蛋白质功能以及开发诊断和治疗剂至关重要。虽然抗体仍然是黄金标准，但它们在动物中的繁琐生产是昂贵的，耗时的，并且可能对许多目标失败。[2]替代技术如单链抗体和适体克服了这些限制中的一些，但仍然需要许多迭代轮的选择和扩增以产生高质量的结合剂。[3]诸如这些的问题已经产生了对下一代技术的迫切需求，这些技术可以作为探索人类蛋白质组的工具，但比现有试剂更有效和更具成本效益。理想的蛋白质捕获试剂将以最小的努力从易于组装的构建模块构建，这些构建模块容易获得且便宜。[4]最有前途的试剂将在已经存在的抗体检测中发挥作用。二价试剂是合成抗体的有吸引力的候选者，因为它们产生的相互作用比相应的单价相互作用强得多。[5]这种方法已被用于创建许多蛋白质亲和试剂，[6]包括最近在体外和细胞中靶向血管内皮生长因子受体2（VEGFR-2）的二价类肽。[7]不幸的是，用于创建这些试剂的发现过程一直难以规模化。最近，我们开发了一种新型的蛋白质亲和试剂，称为“DNA合成体”，其合成不需要体外选择或具有挑战性的合成化学。[8]相反，DNA合成体是通过我们称为核苷酸缀合物（LINC）的配体相互作用的过程产生的。LINC使用短的双链DNA支架来确定将两个配体转化为单个高亲和力蛋白质捕获试剂所需的最佳分离距离和角几何形状。[9]第一章
Creating protein capture reagents on a proteome-wide scale is a grand challenge in molecular medicine.[1] High quality reagents are critical for elucidating protein function, and developing diagnostic and therapeutic agents. Although antibodies remain the gold standard, their cumbersome production in animals is expensive, time consuming, and can fail for many targets.[2] Alternative technologies like single-chain antibodies and aptamers overcome some of these limitations, but still require many iterative rounds of selection and amplification to produce high quality binders.[3] Problems such as these have created a pressing need for next generation technologies that lend themselves as tools to explore the human proteome, but are more efficient and cost-effective to produce than existing reagents. Ideal protein capture reagents would be constructed with minimal effort from easy-toassemble building blocks that are readily available and inexpensive.[4] The most promising reagents would function in assays that already exist for antibodies.Bivalent reagents are attractive candidates for synthetic antibodies because the interactions they produce can be much stronger than their corresponding monovalent interactions.[5] This approach has been used to create many protein affinity reagents,[6] including a recent bivalent peptoid that targets vascular endothelial growth factor receptor 2 (VEGFR-2) in vitro and in cells.[7] Unfortunately, the discovery process used to create these reagents has been difficult to scale. Recently, we developed a new type of protein affinity reagent called a “DNA synbody” whose synthesis does not require in vitro selection or challenging synthetic chemistry.[8] Instead, DNA synbodies are produced by a process that we refer to as ligand interactions by nucleotide conjugates (LINC). LINC uses a short double-stranded DNA scaffold to determine the optimal separation distance and angular geometry needed to transform two ligands into a single high affinity protein capture reagent.[9]
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