1. Fusion to an albumin-binding domain with a high affinity for albumin extends the circulatory half-life and enhances the in vivo antitumor effects of human TRAIL.
- Author
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Li R, Yang H, Jia D, Nie Q, Cai H, Fan Q, Wan L, Li L, and Lu X
- Subjects
- Animals, Antineoplastic Agents administration & dosage, Antineoplastic Agents metabolism, Cell Line, Tumor, Half-Life, Humans, Mice, Inbred BALB C, Mice, Nude, Neoplasms pathology, Protein Binding, Recombinant Fusion Proteins administration & dosage, Recombinant Fusion Proteins metabolism, Recombinant Fusion Proteins pharmacokinetics, Recombinant Fusion Proteins therapeutic use, TNF-Related Apoptosis-Inducing Ligand administration & dosage, TNF-Related Apoptosis-Inducing Ligand metabolism, Antineoplastic Agents pharmacokinetics, Antineoplastic Agents therapeutic use, Neoplasms drug therapy, Serum Albumin metabolism, TNF-Related Apoptosis-Inducing Ligand pharmacokinetics, TNF-Related Apoptosis-Inducing Ligand therapeutic use
- Abstract
Clinical applications of recombinant human tumor necrosis factor-related apoptosis-inducing ligand (hTRAIL) have been limited by their poor pharmacokinetics. Using endogenous albumin as a carrier is an attractive approach for circulatory half-life extension. Here, we produced ABD-hTRAIL and hTRAIL-ABD by fusing the albumin-binding domain (ABD) from protein G to the N- or C-terminus of hTRAIL. We found that ABD-hTRAIL bound human serum albumin (HSA) with a high affinity (0.4 ± 0.18 nM) and formed nanoparticles with an average diameter (~12 nm) above the threshold (~7 nm) of renal filtration. ABD-hTRAIL also bound mouse serum albumin (MSA); thus, its half-life was 40-50-fold greater than that of hTRAIL (14.1 ± 0.87 h vs 0.32 ± 0.14 h). Tumor uptake of ABD-hTRAIL 8-48 h post-injection was 6-16-fold that of hTRAIL. Consequently, the tumor suppression of ABD-hTRAIL in mice bearing subcutaneous xenografts was 3-4 times greater than that of hTRAIL. Additionally, the time period during which ABD-hTRAIL could kill circulating tumor cells was approximately 8 times longer than that of hTRAIL. These results demonstrate that ABD fused to the N-terminus endows hTRAIL with albumin binding ability; once it enters the vasculature, ABD mediates binding with endogenous albumin, thus prolonging the half-life and enhancing the antitumor effect of hTRAIL. However, hTRAIL-ABD did not show a high affinity for albumin and therefore did not display the prolonged circulatory half-life and enhanced antitumor effects. These results demonstrate that N-terminal, but not C-terminal, ABD-fusion is an efficient technique for enhancing the antitumor effects of hTRAIL by using endogenous albumin as a carrier., (Copyright © 2016 Elsevier B.V. All rights reserved.)
- Published
- 2016
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