Evolutionary Qβ Phage Displayed Nanotag Library and Peptides for Biosensing

Author:

Ntemafack Augustin1ORCID,Dzelamonyuy Aristide1ORCID,Nchinda Godwin234ORCID,Bopda Waffo Alain1ORCID

Affiliation:

1. Department of Biochemistry and Molecular Biology, Indiana University School of Medicine, 635 Barnhill Dr., Indianapolis, IN 46202, USA

2. Laboratory of Vaccinology and Biobanking, CIRCB BP 3077 Messa, Yaoundé P.O. Box 3077, Cameroon

3. Department of Pharmaceutical Microbiology and Biotechnology, Nnamdi Azikiwe University, Awka 420110, Nigeria

4. African Center of Excellence for Clinical and Translational Sciences (ACECTS), Yaoundé P.O. Box 13591, Cameroon

Abstract

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.

Funder

National Institutes of Health, National Institute of General Medical Sciences

BIOSENS-Biosensing; National Science Foundation

NIH

Publisher

MDPI AG

Subject

Virology,Infectious Diseases

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