Mäkelä Anna R, Oker-Blom Christian
NanoScience Center, Department of Biological and Environmental Science, P.O. Box 35, FIN-40014 University of Jyväskylä, Finland.
Comb Chem High Throughput Screen. 2008 Feb;11(2):86-98. doi: 10.2174/138620708783744525.
High throughput screening is a core technology in drug discovery. During the past decade, several strategies have been developed to screen (poly)peptide libraries for diverse applications including disease diagnosis and profiling, imaging, as well as therapy. The recently established baculovirus display vector system (BDVS) represents a eukaryotic screening platform that combines the positive attributes of both cell and virus-based display approaches, allowing presentation of complex polypeptides on cellular and viral surfaces. Compared to microbial display systems, the BDVS has the advantage of correct protein folding and post-translational modifications similar to those in mammals, facilitating expression and analysis of proteins with therapeutic interest. The applicability of the system is further expanded by the availability of genetically engineered insect cell lines capable of performing e.g. mammalianized glycosylation in combination with high level of expression. In addition to insect cells, baculovirus can mediate delivery and expression of heterologous genes in a broad spectrum of primary and established mammalian cells. Currently, a variety of baculovirus-based assays aiming at routine high throughput identification of agents targeting cell surface receptors or studies on ligand-receptor interactions are under construction. Here, the advancements and future prospects of the baculovirus display technologies with emphasis on molecular screening and drug delivery applications using insect cell display, mammalian cell display, and virion display are described.
高通量筛选是药物研发中的一项核心技术。在过去十年中，已开发出多种策略来筛选（多）肽文库，用于包括疾病诊断与分析、成像以及治疗等多种应用。最近建立的杆状病毒展示载体系统（BDVS）代表了一种真核筛选平台，它结合了基于细胞和基于病毒的展示方法的优点，能够在细胞和病毒表面呈现复杂的多肽。与微生物展示系统相比，BDVS具有与哺乳动物中相似的正确蛋白质折叠和翻译后修饰的优势，便于表达和分析具有治疗意义的蛋白质。能够进行例如哺乳动物化糖基化并结合高水平表达的基因工程昆虫细胞系的可用性进一步扩展了该系统的适用性。除了昆虫细胞外，杆状病毒还可以介导异源基因在多种原代和已建立的哺乳动物细胞中的递送和表达。目前，正在构建各种基于杆状病毒的检测方法，旨在常规高通量鉴定靶向细胞表面受体的试剂或研究配体 - 受体相互作用。在此，将描述杆状病毒展示技术的进展和未来前景，重点是使用昆虫细胞展示、哺乳动物细胞展示和病毒粒子展示的分子筛选和药物递送应用。
