Rouet Romain, Dudgeon Kip, Christ Daniel
Garvan Institute of Medical Research, Darlinghurst/Sydney, NSW, Australia.
Methods Mol Biol. 2012;907:195-209. doi: 10.1007/978-1-61779-974-7_10.
Human antibody single domains are a promising new class of antibody fragments. Here we describe methods for the cloning of human V(H) and V(L) genes into phage and phagemid vectors. Furthermore, we provide detailed protocols for the generation of single domain antibody libraries by Kunkel mutagenesis and the analysis of diversity by DNA sequencing and superantigen binding.
人源抗体单结构域是一类很有前景的新型抗体片段。在此，我们描述了将人源V(H)和V(L)基因克隆到噬菌体和噬菌粒载体中的方法。此外，我们还提供了通过Kunkel诱变产生单结构域抗体文库以及通过DNA测序和超抗原结合分析多样性的详细方案。
