Division of Life and Pharmaceutical Sciences, The Graduate School, Ewha Womans University, Seoul, 120-750, Korea.
Mol Cells. 2011 Jun;31(6):509-13. doi: 10.1007/s10059-011-2268-8. Epub 2011 Apr 15.
Target-specific antibodies can be rapidly enriched and identified from an antibody library using phage display. Large, naïve antibody libraries derived from synthetic or unimmunized sources can yield antibodies against virtually any antigen, whereas libraries from immunized sources tend to be smaller and are used exclusively against the antigen of immunization. In this study, 25 scFv libraries made from the spleens of immunized rabbits, each with a size ranging from 10(8) to higher than 10(9), were combined into a single large library with > 10(10) individual clones. Panning of this combined library yielded target-specific rabbit scFv clones against many non-immunizing antigens, including proteins, peptides, and a small molecule. Notably, specific scFv clones against a rabbit self-antigen (rabbit serum albumin) and a phosphorylated protein (epidermal growth factor receptor pTyr1173) could be isolated from the library. These results suggest that the immune library contained a significant number of unimmunized clones and that a sufficiently large immune library can be utilized similarly to a naïve library, i.e., against various non-immunizing antigens to yield specific antibodies.
利用噬菌体展示技术，可以从抗体文库中快速富集和鉴定针对特定靶标的抗体。从合成或未免疫来源衍生的大型、天然抗体文库可以产生针对几乎任何抗原的抗体，而来自免疫来源的文库通常较小，专门用于免疫的抗原。在这项研究中，将来自免疫兔脾脏的 25 个 scFv 文库合并成一个单一的大型文库，每个文库的大小范围从 10(8)到 10(9)以上，文库的大小范围从 10(8)到 10(9)以上。对这个组合文库进行淘选，得到了针对许多非免疫原性抗原的针对特定靶标的兔 scFv 克隆，包括蛋白质、肽和小分子。值得注意的是，能够从文库中分离出针对兔自身抗原（兔血清白蛋白）和磷酸化蛋白（表皮生长因子受体 pTyr1173）的特异性 scFv 克隆。这些结果表明，免疫文库中包含大量未免疫的克隆，并且足够大的免疫文库可以像天然文库一样被利用，即针对各种非免疫原性抗原产生特异性抗体。
