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Multivalent Peptide Dendrimers for Targeting

机译：用于靶向的多价肽树枝状大分子

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Phage display is a widely used technique in molecular biology for the identification of selective, peptide-based ligands for a desired proteinogenic substrate. Biological combinatorial libraries of the M13 bacteriophage are commonly used for this purpose (Figure 1). The M13 bacteriophage consists of a single-stranded circular piece of DNA (～6400 bases) encapsulated by 2700 copies of the pVIII major capsid protein. At the terminus of this assembly is presented five copies of the targeting peptide, usually 7-15 aa, fused at their C-terminus to pIII. The presented peptides can be randomized (or focused) at the DNA level, and the respective phage constituted in a library, e.g. ～10~9 unique members in a library consisting of phage with 7-mer peptides. When introduced to a substrate, in vivo or in vitro, members from this library may engage in substrate binding. Phage that bind to the substrate are then amplified in E. coli and reconstituted into libraries that are consequently enriched with phage that bind to the substrate. After additional rounds of selection and amplification, individual clones may be isolated and routine DNA sequencing can be used to identify the exact peptide sequence responsible. Interestingly, individual phage peptides often show only weak binding to their intended substrate (K_d ～ 100 – 1000 μM) while the phage themselves have K_d values in the nanomolar range. This superior affinity is due to the inherent, multivalent character of the phage and its association with substrates through multiple weak interactions. Nevertheless, most ligand-directed targeting strategies involving phage peptides rely on the conjugation of a single phage peptide to the active payload.
机译：噬菌体显示是用于鉴定所需蛋白原基质的选择性，肽基配体的分子生物学中的广泛使用的技术。 M13噬菌体的生物组合文库通常用于此目的（图1）。 M13噬菌体由单链圆形的DNA（〜6400个碱基）组成，其包封的PVIII主要衣壳蛋白的2700份。在该组件的末端，呈现靶向肽的五个拷贝，通常为7-15AA，在其C-末端融合给PIII。所呈现的肽可以在DNA水平随机化（或聚焦），以及在文库中构成的各个噬菌体，例如， 〜10〜9个独特的成员在由具有7-MEL肽的噬菌体组成的文库中。当引入底物时，在体内或体内，该文库的构件可以与底物结合接合。然后在大肠杆菌中扩增与基质结合的噬菌体，并重构成后来富含与基材结合的噬菌体的文库。在额外的选择和扩增之后，可以分离各个克隆，并且可以使用常规DNA测序来鉴定负责的精确肽序列。有趣的是，单个噬菌体肽通常仅显示与其预期基质（K_D〜100-1000μm）的弱结合，而噬菌体本身在纳米摩尔范围内具有K_D值。这种优异的亲和力是由于噬菌体的固有，多价特征及其通过多个弱相互作用与基材的关系。然而，涉及噬菌体肽的大多数配体定向靶向策略依赖于单噬菌体肽的缀合到活性有效载荷。

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