重组DNA
噬菌体展示
生物
计算生物学
抗体
分子生物学
单域抗体
癌症免疫疗法
克隆(编程)
重链
免疫疗法
癌症
遗传学
基因
计算机科学
程序设计语言
作者
Damjana Kastelic,Snjez̆ana Frković‐Grazio,Daniel Baty,Gilles Truan,Radovan Komel,Denis Pompon
标识
DOI:10.1016/j.jim.2009.08.016
摘要
Heavy chain antibodies are naturally occurring in camelidae (camels and llamas). Their variable domain (VHH) can be efficiently produced as a recombinant protein in E. coli with a large range of applications in the fields of diagnostics and immunotherapy. Standard cloning approach involves resolution of VHH from the heavy chain variable domain of conventional antibodies (VH) by a nested PCR amplification followed by a phage display based selection. Present work illustrates that in contrast to usual finding, specific, good affinity and efficiently expressed VH domain of conventional antibodies can be selected from the co-amplification products of VH and VHH cDNAs. Sequence analysis illustrated that following the two first rounds of selection against cancer markers, similar number of VH and VHH binders were observed. However, after a third round, the more specific binders directed against p53, VEGF, BCL-2 proteins surprisingly contain only VH specific hallmarks. Characterisation of the specificity, affinity and productivity of selected VH binders is described. Because llama VHs show higher sequence and structural homology with the human VH III group than llama VHHs (Vu et al., 1997), they constitute very interesting agents in therapeutic applications, especially in human immunotherapy and cancer treatment.
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