化学
变性(裂变材料)
重组DNA
片段(逻辑)
免疫球蛋白Fab片段
生物物理学
免疫球蛋白Fc片段
领域(数学分析)
结晶学
抗体
理论(学习稳定性)
立体化学
生物化学
免疫球蛋白G
肽序列
互补决定区
生物
计算机科学
数学
遗传学
基因
机器学习
数学分析
程序设计语言
核化学
作者
Daniela Röthlisberger,Annemarie Honegger,Andreas Plückthun
标识
DOI:10.1016/j.jmb.2005.01.053
摘要
Recombinant antibody fragments, most notably Fab and scFv, have become important tools in research, diagnostics and therapy. Since different recombinant antibody formats exist, it is crucial to understand the difference in their respective biophysical properties. We assessed the potential stability benefits of changing the scFv into the Fab format, the influence of the variable domains on the stability of the Fab fragment, and the influence of the interchain disulfide bond in the Fab fragment. To analyze domain interactions, the Fab fragment was broken down into its individual domains, several two-domain assemblies and one three-domain assembly. The equilibrium denaturation properties of these constructs were then compared to those of the Fab fragment. It was found that mutual stabilization occurred across the VH/VL and the CH1/CL interface, whereas the direct interaction between the VL and the CL domain had no influence on the stability of either domain. This observation can be explained by the different interfaces used for interaction. In contrast, the whole CH1CL and VHVL unit showed significant mutual stabilization, indicating a high degree of cooperation between the VH/VL and CH1/CL interface. The interchain disulfide bond in the Fab fragment plays an essential role in this stabilization. In addition to the effects of domain association on the thermodynamic (equilibrium) stability, Fab fragments differ from scFv fragments of similar equilibrium stability by having a very slow unfolding rate. This kinetic stabilization may increase significantly the resistance of Fab fragments against short time exposure to adverse conditions.
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