PT - JOURNAL ARTICLE AU - Zhuolun Shen AU - Yufei Xiang AU - Sandra Vegara AU - Apeng Chen AU - Zhengyun Xiao AU - Ulises Santiago AU - Changzhong Jin AU - Zhe Sang AU - Jiadi Luo AU - Kong Chen AU - Dina Schneidman-Duhovny AU - Carlos Camacho AU - Guillermo Calero AU - Baoli Hu AU - Yi Shi TI - A robust and versatile nanobody platform for drug delivery AID - 10.1101/2020.08.19.257725 DP - 2020 Jan 01 TA - bioRxiv PG - 2020.08.19.257725 4099 - http://biorxiv.org/content/early/2020/08/20/2020.08.19.257725.1.short 4100 - http://biorxiv.org/content/early/2020/08/20/2020.08.19.257725.1.full AB - Therapeutic and diagnostic efficacies of numerous small biomolecules and chemical compounds are hampered by the short half-lives. Here we report the development of a repertoire of diverse, high-affinity albumin-nanobodies (NbHSA) to facilitate drug delivery. By integrating biophysics, and hybrid structural biology, we have systematically characterized the NbHSA for albumin binding, mapped the epitopes, and resolved the architecture of a tetrameric Nb-albumin complex. We employed quantitative proteomics for accurate, multiplex Nb pharmacokinetic analysis. Using a humanized albumin mouse model, we found that the NbHSA has outstanding pharmacokinetics; the most stable NbHSA has a 771-fold T1/2 improvement compared with a control Nb. Interestingly, the pharmacokinetics of NbHSA is related to their biophysical and structural properties. To demonstrate the utility of NbHSA, we developed a highly stable NbHSA-hIL-2 cytokine conjugate “Duraleukin” and confirmed its improved anticancer properties than hIL-2 alone. We envision that this high-quality Nb resource will advance research into novel biotherapeutics.Competing Interest StatementThe University of Pittsburgh has filed a provisional patent encompassing the technologies described in this manuscript.