期刊
BIOCONJUGATE CHEMISTRY
卷 33, 期 3, 页码 452-462出版社
AMER CHEMICAL SOC
DOI: 10.1021/acs.bioconjchem.1c00579
关键词
-
类别
资金
- National Science Foundation [1510817, 1144726]
- NIH-NIGMS [P20 GM103445]
- NSF [IIA-1301765]
- Direct For Education and Human Resources
- Division Of Graduate Education [1144726] Funding Source: National Science Foundation
- Div Of Chem, Bioeng, Env, & Transp Sys
- Directorate For Engineering [1510817] Funding Source: National Science Foundation
In this study, SpyTag peptide was fused to the surface of engineered E2 nanocages to enable tunable nanocage decoration and effective E2 cell targeting using a variety of SpyCatcher fusion proteins. The core of the E2 nanocage incorporated four phenylalanine mutations for hydrophobic drug loading and pH-responsive release. By functionalizing the nanocage surface with a highly cell-specific EGFR-targeting protein conjugate, efficient drug loading and targeted cell death in inflammatory breast cancer cells were achieved. This engineered protein nanocage construct demonstrates versatility for cargo delivery.
Naturally occurring protein nanocages are promising drug carriers because of their uniform size and biocompatibility. Engineering efforts have enhanced the delivery properties of nanocages, but cell specificity and high drug loading remain major challenges. Herein, we fused the SpyTag peptide to the surface of engineered E2 nanocages to enable tunable nanocage decoration and effective E2 cell targeting using a variety of SpyCatcher (SC) fusion proteins. Additionally, the core of the E2 nanocage incorporated four phenylalanine mutations previously shown to allow hydrophobic loading of doxorubicin and pH-responsive release in acidic environments. We functionalized the surface of the nanocage with a highly cell-specific epidermal growth factor receptor ( EGFR)-targeting protein conjugate, 4GE11-mCherry-SC, developed previously in our laboratories by employing unnatural amino acid (UAA) protein engineering chemistries. Herein, we demonstrated the benefits of this engineered protein nanocage construct for efficient drug loading, with a straightforward method for removal of the unloaded drug through elastin-like polypeptide-mediated inverse transition cycling. Additionally, we demonstrated approximately 3-fold higher doxorubicin internalization in inflammatory breast cancer cells compared to healthy breast epithelial cells, leading to targeted cell death at concentrations below the ICSO of free doxorubicin. Collectively, these results demonstrated the versatility of our UAA-based EGFR-targeting protein construct to deliver a variety of cargoes efficiently, including engineered E2 nanocages capable of site-specific functionalization and doxorubicin loading.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据