Cekaite Lina, Hovig Eiving, Sioud Mouldy
Department of Immunology, Institute for Cancer Research, The Norwegian Radium Hospital, Rikshopitalet University Hospital Montebello, N-0310 Oslo, Norway.
Methods Mol Biol. 2009;524:273-85. doi: 10.1007/978-1-59745-450-6_20.
Successful adaptation of microarray technology for high-throughput screening of proteins requires a large number of purified recombinant proteins, e.g., antibodies for use as capture molecules. Phage surface display technology has been used for the surface expression of proteins, peptides or cDNA repertoires expressed by tumor cells. It does not require protein purification, as recombinant phages can be spotted on glass slides and used in a high-throughput screening format. Biopanning of phage libraries on patient serum antibodies is expected to enrich for antibody-binding phages for the fabrication of diagnostic and/or prognostic B-cell epitope microarrays. In contrast to other immunological techniques, microarrays can measure the antibody levels against different epitopes in a single test. This chapter highlights the recent advances in phage-based microarray technology to profile humoral immune responses in cancer patients.
要成功将微阵列技术应用于蛋白质的高通量筛选，需要大量纯化的重组蛋白，例如用作捕获分子的抗体。噬菌体表面展示技术已用于肿瘤细胞表达的蛋白质、肽或cDNA文库的表面表达。由于重组噬菌体可以点样在载玻片上并用于高通量筛选，因此无需进行蛋白质纯化。利用患者血清抗体对噬菌体文库进行生物淘选，有望富集与抗体结合的噬菌体，用于制备诊断和/或预后B细胞表位微阵列。与其他免疫技术相比，微阵列可以在一次检测中测量针对不同表位的抗体水平。本章重点介绍基于噬菌体的微阵列技术在分析癌症患者体液免疫反应方面的最新进展。
