通过一步克隆构建大型天然人噬菌体展示Fab文库。
Construction of a large naïve human phage-displayed Fab library through one-step cloning.
作者信息
Zhu Zhongyu, Dimitrov Dimiter S
机构信息
NCI-Frederick, National Institutes of Health, Frederick, MD, USA.
出版信息
Methods Mol Biol. 2009;525:129-42, xv. doi: 10.1007/978-1-59745-554-1_6.
Abstract
Antibody-based therapeutics is attracting more attention in the post-genome era, in contrast to a diminution in the initial high expectation for rapid development of gene-based therapeutic modalities. In support to the antibody-based therapeutics, the advent of recent technologies has made human antibody screening and production progressively more economic. Among those technologies, phage-display antibody library has been successfully applied in the antibody-based drug development both as fully human antibody sources and tools for antibody engineering. Building up a high-quality antibody library with a large library size and high diversity has been crucial for successful isolation of antibodies. Here we describe an efficient strategy for the construction of a large naïve phage-display human Fab library with one-step cloning. Optimization of each key step is extensively discussed and simplified protocols for library panning and Fab production are also described.
摘要
在后基因组时代，基于抗体的治疗方法正吸引着更多关注，这与人们最初对基于基因的治疗方法快速发展的高度期望有所下降形成对比。为支持基于抗体的治疗方法，近期技术的出现使人类抗体筛选和生产逐渐变得更加经济。在这些技术中，噬菌体展示抗体库已成功应用于基于抗体的药物开发，既作为全人源抗体来源，也作为抗体工程工具。构建具有大库容量和高多样性的高质量抗体库对于成功分离抗体至关重要。在此，我们描述了一种通过一步克隆构建大型天然噬菌体展示人源Fab库的有效策略。文中广泛讨论了每个关键步骤的优化，并描述了用于库筛选和Fab生产的简化方案。
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