Department of Protein Science, KTH - Royal Institute of Technology, Stockholm, Sweden.
Methods Mol Biol. 2023;2681:99-112. doi: 10.1007/978-1-0716-3279-6_7.
This review describes the principles for generation of affibody molecules using bacterial display on the Gram-negative Escherichia coli and the Gram-positive Staphylococcus carnosus, respectively. Affibody molecules are small and robust alternative scaffold proteins that have been explored for therapeutic, diagnostic, and biotechnological applications. They typically exhibit high-stability, affinity, and specificity with high modularity of functional domains. Due to the small size of the scaffold, affibody molecules are rapidly excreted through renal filtration and can efficiently extravasate from blood and penetrate tissues. Preclinical and clinical studies have demonstrated that affibody molecules are promising and safe complements to antibodies for in vivo diagnostic imaging and therapy. Sorting of affibody libraries displayed on bacteria using fluorescence-activated cell sorting is an effective and straightforward methodology and has been used successfully to generate novel affibody molecules with high affinity for a diverse range of molecular targets.
本文综述了分别在革兰氏阴性的大肠杆菌和革兰氏阳性的肉葡萄球菌上利用细菌展示来生成亲和体分子的原理。亲和体分子是体积小且稳定的替代支架蛋白,已被探索用于治疗、诊断和生物技术应用。它们通常表现出高稳定性、高亲和力和特异性,并且具有多功能域的高模块性。由于支架较小,亲和体分子可通过肾脏过滤迅速排泄,并能有效地从血液中外渗并穿透组织。临床前和临床研究表明,亲和体分子是抗体在体内诊断成像和治疗方面有前途且安全的补充。使用荧光激活细胞分选对细菌上展示的亲和体文库进行分类是一种有效且直接的方法,已成功用于生成针对多种分子靶标具有高亲和力的新型亲和体分子。