Robin Gautier, Martineau Pierre
BioXtal SA, Marseille, France.
Methods Mol Biol. 2012;907:109-22. doi: 10.1007/978-1-61779-974-7_6.
Libraries of antibody fragments displayed on filamentous phages have proved their value to generate human antibodies against virtually any target. We describe here a simple protocol to make large and diverse libraries based on a single or few frameworks. Diversity is introduced in the third hypervariable loops using degenerate synthetic oligonucleotides and PCR assembly. Because all the antibody fragments isolated from the library will share their framework sequence, their stability and physical properties will be more consistent and customizable than when antibody fragments are isolated from a library prepared from human donors.
展示在丝状噬菌体上的抗体片段文库已证明其在生成针对几乎任何靶标的人源抗体方面的价值。我们在此描述一种简单方案，以基于单个或少数几个框架构建大型且多样的文库。使用简并合成寡核苷酸和PCR组装在第三个高变环中引入多样性。由于从文库中分离出的所有抗体片段将共享其框架序列，与从人类供体制备的文库中分离抗体片段相比，它们的稳定性和物理性质将更加一致且可定制。
