Evolutionary Qβ Phage Displayed Nanotag Library and Peptides for Biosensing.

Evolutionary Qβ Phage Displayed Nanotag Library and Peptides for Biosensing.
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DOI:
10.3390/v15071414
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发表时间:
2023-06-22
期刊:
Viruses
影响因子:
--
通讯作者:
Bopda Waffo A
Bopda Waffo A
中科院分区:
其他
文献类型:
--
作者:
Ntemafack A;Dzelamonyuy A;Nchinda G;Bopda Waffo A

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我们选择了一种新的生物素结合肽传感生物素,生物素化的蛋白质,和核苷酸。从展示在RNA大肠杆菌噬菌体Qβ上的15-mer文库中,鉴定了15-氨基酸长的肽(HGHGWQIPVWPWGQG),其在此被称为纳米标签以选择性地结合生物素。通过感染洗脱淘选实现靶选择。在Qβ噬菌体平台上,将筛选出的肽段作为口蹄疫病毒(FMDV)免疫原性表位的转换子进行测试。生物素-标签显示对表位对其同源抗体(SD 6)的亲和力没有显著影响。纳米标签结合的生物素选择性地融合到表位的C-或N-末端。当位于纳米标签的N末端时,表位将不结合或识别SD 6。此外,在竞争性ELISA中,生物素与SD 6抗体线性竞争。使用所选择的重组噬菌体本身作为探针或换能器的竞争测定使得该技术能够作为生物传感器工具包来感测和定量SD 6分析物。在本文中,已发表的Strep II纳米标签(DVEWLDERVPLVET)被用作对照,并且具有与我们提出的新型生物素标签相似的功能,从而为开发用于诊断目的的设备提供了新的平台。
We selected a novel biotin-binding peptide for sensing biotin, biotinylated proteins, and nucleotides. From a 15-mer library displayed on the RNA coliphage Qβ, a 15-amino acid long peptide (HGHGWQIPVWPWGQG) hereby referred to as a nanotag was identified to selectively bind biotin. The target selection was achieved through panning with elution by infection. The selected peptide was tested as a transducer for an immunogenic epitope of the foot-and-mouth disease virus (FMDV) on Qβ phage platform separated by a linker. The biotin-tag showed no significant influence on the affinity of the epitope to its cognate antibody (SD6). The nanotag-bound biotin selectively fused either to the C- or N-terminus of the epitope. The epitope would not bind or recognize SD6 while positioned at the N-terminus of the nanotag. Additionally, the biotin competed linearly with the SD6 antibody in a competitive ELISA. Competition assays using the selected recombinant phage itself as a probe or transducer enable the operationalization of this technology as a biosensor toolkit to sense and quantify SD6 analyte. Herein, the published Strep II nanotag (DVEWLDERVPLVET) was used as a control and has similar functionalities to our proposed novel biotin-tag thereby providing a new platform for developing devices for diagnostic purposes.
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