A resource of high-quality and versatile nanobodies for drug delivery
Zhuolun Shen,
Yufei Xiang,
Sandra Vergara,
Apeng Chen,
Zhengyun Xiao,
Ulises Santiago,
Changzhong Jin,
Zhe Sang,
Jiadi Luo,
Kong Chen,
Dina Schneidman-Duhovny,
Carlos Camacho,
Guillermo Calero,
Baoli Hu,
Yi Shi
Affiliations
Zhuolun Shen
Department of Cell Biology, University of Pittsburgh, Pittsburgh, PA, USA; School of Medicine, Tsinghua University, Beijing, China
Yufei Xiang
Department of Cell Biology, University of Pittsburgh, Pittsburgh, PA, USA
Sandra Vergara
Department of Structural Biology, University of Pittsburgh, Pittsburgh, PA, USA
Apeng Chen
Department of Neurological Surgery, University of Pittsburgh, Pittsburgh, PA, USA; Pediatric Neurosurgery, UPMC Children's Hospital of Pittsburgh, Pittsburgh, PA, USA
Zhengyun Xiao
Department of Cell Biology, University of Pittsburgh, Pittsburgh, PA, USA
Ulises Santiago
Department of Computational and Systems Biology, University of Pittsburgh, Pittsburgh, PA, USA
Changzhong Jin
Department of Cell Biology, University of Pittsburgh, Pittsburgh, PA, USA
Zhe Sang
Department of Cell Biology, University of Pittsburgh, Pittsburgh, PA, USA; University of Pittsburgh-Carnegie Mellon University Joint Program for Computational Biology, Pittsburgh, PA, USA
Jiadi Luo
Department of Medicine, University of Pittsburgh, Pittsburgh, PA, USA
Kong Chen
Department of Medicine, University of Pittsburgh, Pittsburgh, PA, USA
Dina Schneidman-Duhovny
School of Computer Science and Engineering, Institute of Life Sciences, University of Jerusalem, Tambaram, Israel
Carlos Camacho
Department of Computational and Systems Biology, University of Pittsburgh, Pittsburgh, PA, USA
Guillermo Calero
Department of Structural Biology, University of Pittsburgh, Pittsburgh, PA, USA
Baoli Hu
Department of Neurological Surgery, University of Pittsburgh, Pittsburgh, PA, USA; Pediatric Neurosurgery, UPMC Children's Hospital of Pittsburgh, Pittsburgh, PA, USA; Molecular and Cellular Cancer Biology Program, UPMC Hillman Cancer Center, Pittsburgh, PA, USA
Yi Shi
Department of Cell Biology, University of Pittsburgh, Pittsburgh, PA, USA; University of Pittsburgh-Carnegie Mellon University Joint Program for Computational Biology, Pittsburgh, PA, USA; Corresponding author
Summary: 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.
