单个寻址的单链Fv抗体分子的抗原结合力。
Antigen binding forces of individually addressed single-chain Fv antibody molecules.
作者信息
Ros R, Schwesinger F, Anselmetti D, Kubon M, Schäfer R, Plückthun A, Tiefenauer L
机构信息
Paul Scherrer Institut, CH-5232 Villigen PSI, Switzerland.
出版信息
Proc Natl Acad Sci U S A. 1998 Jun 23;95(13):7402-5. doi: 10.1073/pnas.95.13.7402.
Abstract
Antibody single-chain Fv fragment (scFv) molecules that are specific for fluorescein have been engineered with a C-terminal cysteine for a directed immobilization on a flat gold surface. Individual scFv molecules can be identified by atomic force microscopy. For selected molecules the antigen binding forces are then determined by using a tip modified with covalently immobilized antigen. An scFv mutant of 12% lower free energy for ligand binding exhibits a statistically significant 20% lower binding force. This strategy of covalent immobilization and measuring well separated single molecules allows the characterization of ligand binding forces in molecular repertoires at the single molecule level and will provide a deeper insight into biorecognition processes.
摘要
已设计出对荧光素具有特异性的抗体单链Fv片段（scFv）分子，其C末端带有半胱氨酸，用于定向固定在平坦的金表面上。单个scFv分子可通过原子力显微镜识别。对于选定的分子，然后使用共价固定抗原修饰的探针来确定抗原结合力。配体结合自由能降低12%的scFv突变体表现出具有统计学意义的20%的较低结合力。这种共价固定和测量分离良好的单分子的策略能够在单分子水平上表征分子库中的配体结合力，并将为生物识别过程提供更深入的见解。
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