Isolation of Cowpea Mosaic Virus-Binding Peptides.
Isolation of Cowpea Mosaic Virus-Binding Peptides.
复制标题
豇豆花叶病毒结合肽的分离
DOI：
10.1021/acs.biomac.1c00712
复制
发表时间：
2021-08-09
影响因子：
6.2
通讯作者：
Steinmetz, Nicole F.
中科院分区：
文献类型：
--
作者：
Chan, Soo Khim;Steinmetz, Nicole F.
植物病毒豇豆花叶病毒（CPMV）是一种天然的纳米载体，已开发为用于递送各种有效载荷的平台技术，包括用于疫苗的肽表位、用于成像的造影剂和用于治疗的药物。基因融合和化学缀合是将活性成分负载到CPMV外部和/或内部的主要方法。然而，这些方法有局限性;基因工程仅限于生物制剂，化学改变通常需要多步反应，同时修饰CPMV和活性成分。这两种方法都可能导致粒子不稳定。因此，为了提供一个替代的途径CPMV功能化，我们报告的肽，特异性结合到CPMV，称为CPMV结合肽（CBP）的分离。我们使用商业M13噬菌体展示7-mer肽文库来淘选和选择选择性结合CPMV的肽。先导候选物的生物淘选和表征导致基序“GWRVSEF/L”作为CPMV特异性基序的分离，其中在第七位置处的苯丙氨酸（F）比亮氨酸（L）强。证明了对CPMV的特异性，未观察到对其他植物病毒的交叉反应性。为了证明货物装载，GWRVSEF用生物素、异硫氰酸荧光素（FITC）和人表皮生长因子受体2（HER 2）特异性靶向肽配体标记。证实了活性成分的展示，并证明了标记的和靶向的CPMV在细胞结合测定中的效用。CBP功能化策略为CPMV纳米颗粒功能化提供了一种新的途径，并且应该提供一种通用的工具来添加否则可能难以缀合或展示的活性成分。
The plant virus cowpea mosaic virus (CPMV) is a natural nanocarrier that has been developed as a platform technology for the delivery of various payloads including peptide epitopes for vaccines, contrast agents for imaging, and drugs for therapy. Genetic fusion and chemical conjugations are the mainstay approaches to load the active ingredient to the exterior and/or interior of CPMV. However, these methods have limitations; genetic engineering is limited to biologics, and chemical alteration often requires multistep reactions with modification of both CPMV and the active ingredient. Either method can also result in particle instability. Therefore, to provide an alternate path toward CPMV functionalization, we report the isolation of peptides that specifically bind to CPMV, termed CPMV-binding peptides (CBP). We used a commercial M13 phage display 7-mer peptide library to pan for and select peptides that selectively bind to CPMV. Biopanning and characterization of lead candidates resulted in isolation of the motif “GWRVSEF/L” as the CPMV-specific motif with phenylalanine (F) at the seventh position being stronger than leucine (L). Specificity to CPMV was demonstrated, and cross-reactivity toward other plant viruses was not observed. To demonstrate cargo loading, GWRVSEF was tagged with biotin, fluorescein isothiocyanate (FITC), and a human epidermal growth factor receptor 2 (HER2)-specific targeting peptide ligand. Display of the active ingredient was confirmed, and utility of tagged and targeted CPMV in cell binding assays was demonstrated. The CBP functionalization strategy offers a new avenue for CPMV nanoparticle functionalization and should offer a versatile tool to add active ingredients that otherwise may be difficult to conjugate or display.
登录
查看更多内容
影响因子：
8
作者：
Lomonossoff GP
通讯作者：
Lomonossoff GP
影响因子：
3.1
作者：
Madi, Mikidache;Mioulet, Valerie;King, Donald P.;Lomonossoff, George P.;Montague, Nicholas P.
通讯作者：
Montague, Nicholas P.
影响因子：
3.7
作者：
Nkanga CI;Steinmetz NF
通讯作者：
Steinmetz NF
影响因子：
4.6
作者：
Qiang X;Sun K;Xing L;Xu Y;Wang H;Zhou Z;Zhang J;Zhang F;Caliskan B;Wang M;Qiu Z
通讯作者：
Qiu Z
影响因子：
4.6
作者：
Wang, Chao;Fiering, Steven N.;Steinmetz, Nicole F.
通讯作者：
Steinmetz, Nicole F.