Saerens Dirk, Huang Lieven, Bonroy Kristien, Muyldermans Serge
Laboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Pleinlaan 2, B-1050 Brussels, Belgium.
Department of Molecular and Cellular Interactions, VIB, Brussels, Belgium.
Sensors (Basel). 2008 Aug 8;8(8):4669-4686. doi: 10.3390/s8084669.
Today's proteomic analyses are generating increasing numbers of biomarkers, making it essential to possess highly specific probes able to recognize those targets. Antibodies are considered to be the first choice as molecular recognition units due to their target specificity and affinity, which make them excellent probes in biosensor development. However several problems such as difficult directional immobilization, unstable behavior, loss of specificity and steric hindrance, may arise from using these large molecules. Luckily, protein engineering techniques offer designed antibody formats suitable for biomarker analysis. Minimization strategies of antibodies into Fab fragments, scFv or even single-domain antibody fragments like VH, VL or VHHs are reviewed. Not only the size of the probe but also other issues like choice of immobilization tag, type of solid support and probe stability are of critical importance in assay development for biosensing. In this respect, multiple approaches to specifically orient and couple antibody fragments in a generic one-step procedure directly on a biosensor substrate are discussed.
如今的蛋白质组学分析正在产生越来越多的生物标志物，因此拥有能够识别这些靶标的高特异性探针至关重要。由于抗体的靶标特异性和亲和力，它们被认为是作为分子识别单元的首选，这使得它们成为生物传感器开发中的优秀探针。然而，使用这些大分子可能会出现一些问题，如定向固定困难、行为不稳定、特异性丧失和空间位阻。幸运的是，蛋白质工程技术提供了适用于生物标志物分析的设计抗体形式。本文综述了将抗体最小化为Fab片段、单链抗体片段（scFv）甚至单域抗体片段（如VH、VL或VHH）的策略。在生物传感检测开发中，不仅探针的大小，而且其他问题，如固定标签的选择、固体支持物的类型和探针稳定性，都至关重要。在这方面，本文讨论了在通用的一步法中直接在生物传感器基质上特异性定向和偶联抗体片段的多种方法。
