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A resource of high-quality and versatile nanobodies for drug delivery

2021; Cell Press; Volume: 24; Issue: 9 Linguagem: Inglês

10.1016/j.isci.2021.103014

2589-0042

Zhuolun Shen, Yufei Xiang, Sandra Vergara, Apeng Chen, Zhengyun Xiao, Ulises Santiago, Changzhong Jin, Zhe Sang, Jiadi Luo, Kong Chen, Dina Schneidman‐Duhovny, Carlos J. Camacho, Guillermo Calero, Baoli Hu, Yi Shi,

Protein purification and stability

Therapeutic and diagnostic efficacies of small biomolecules and chemical compounds are hampered by suboptimal pharmacokinetics. Here, we developed a repertoire of robust and high-affinity antihuman serum albumin nanobodies (NbHSA) that can be readily fused to small biologics for half-life extension. We characterized the thermostability, binding kinetics, and cross-species reactivity of NbHSAs, mapped their epitopes, and structurally resolved a tetrameric HSA-Nb complex. We parallelly determined the half-lives of a cohort of selected NbHSAs in an HSA mouse model by quantitative proteomics. Compared to short-lived control nanobodies, the half-lives of NbHSAs were drastically prolonged by 771-fold. NbHSAs have distinct and diverse pharmacokinetics, positively correlating with their albumin binding affinities at the endosomal pH. We then generated stable and highly bioactive NbHSA-cytokine fusion constructs "Duraleukin" and demonstrated Duraleukin's high preclinical efficacy for cancer treatment in a melanoma model. This high-quality and versatile Nb toolkit will help tailor drug half-life to specific medical needs.