Department of Chemistry, Materials and Chemical Engineering Giulio Natta, Politecnico di Milano, Via Mancinelli 7, 20131 Milano, Italy.
Curr Pharm Biotechnol. 2011 Oct;12(10):1537-47. doi: 10.2174/138920111798357285.
The technique of combinatorial peptide ligand libraries (CPLL), for capturing and amplifying low-abundance proteins in r-DNA products as well as in a number of other biological systems, is here analyzed in depth and reviewed. This methodology is based on a creation of several millions of bio-specific ligands composed of hexapeptides produced in a combinatorial way. When acting on an overloading and saturation principle, high-abundance species are captured in limited amounts, whereas low-abundance ones keep being concentrated on their bio-specific ligand till substantial harvesting from solution (the capture process occurring in general from ca. 50% up to 90% efficiency). Examples are given on tracking host-cell impurities present in, e.g., recombinant albumin or monoclonal antibodies. Additionally, other examples of detecting traces of additives and fining agents in such beverages as white and red wines are presented. The unique mechanisms underlying the protein capture in the CPLL methodology, as opposed to capture by homogeneous beads, as represented by ion-exchangers and by hydrophobic resins, are discussed in depth.
组合肽配体文库 (CPLL) 技术用于捕获和扩增 r-DNA 产物以及许多其他生物系统中的低丰度蛋白质，本文深入分析和回顾了该方法。该方法基于创建数百万种由组合方式产生的六肽组成的生物特异性配体。当采用超载和饱和原理时，高丰度物质的捕获量有限，而低丰度物质则继续在其生物特异性配体上浓缩，直到从溶液中大量收获（捕获过程通常效率为 50%至 90%）。例如，跟踪重组白蛋白或单克隆抗体中存在的宿主细胞杂质。此外，还介绍了在白葡萄酒和红葡萄酒等饮料中检测添加剂和澄清剂痕迹的其他例子。本文深入讨论了 CPLL 方法中蛋白质捕获的独特机制，与通过离子交换剂和疏水树脂等均相珠粒进行捕获的机制相反。
