Evolutionary Qβ Phage Displayed Nanotag Library and Peptides for Biosensing

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

Mapping immunological and host receptor binding determinants of SARS-CoV spike protein utilizing the Qubevirus platform

The motifs involved in tropism and immunological interactions of SARS-CoV spike (S) protein were investigated utilizing the Qubevirus platform. We showed that separately, 14 overlapping peptide fragments representing the S protein (F1-14 of 100 residues each) could be inserted into the C terminus of A1 on recombinant Qubevirus without affecting its viability. Additionally, recombinant phage expression resulted in the surface exposure of different engineered fragments in an accessible manner. The F6 from S425-525 was found to contain the binding determinant of the recombinant human angiotensin-converting enzyme 2, with the shortest active binding motif situated between residues S437-492. Upstream, another fragment, F7, containing an overlapping portion of F6 would not bind to recombinant human angiotensin-converting enzyme 2, confirming that a contiguous stretch of residues could adopt the appropriate structural orientation of F6 as an insertion within the Qubevirus. The F6 (S441-460) and other inserts, including F7/F8 (S601-620) and F10 (S781-800), were demonstrated to contain important immunological determinants through recognition and binding of S protein specific (anti-S) antibodies. An engineered chimeric insert bearing the fusion of all three anti-S reactive epitopes improved substantially the recognition and binding to their cognate antibodies. These results provide insights into humoral immune relevant epitopes and tropism characteristics of the S protein with implications for the development of subunit vaccines or other biologics against SARS-CoV

Application of heat-treated oyster shell powder to induce priming of Theobroma cocoa seedlings plant defense system against Phytophthora megakarya attack

Priming with natural compounds is an interesting alternative for sustainable agriculture to explore the molecular mechanisms associated with the management of diseases caused by soil borne pathogens. The aim of this work was to evaluate the effect of the soil amendment with bio-oyster shell on the stimulation of the cocoa plant defense systems against P. megakarya in the nursery. For this purpose, the potential of bio-oyster shell to stimulate cocoa plant defense mechanisms was evaluated by soil amendment at 1% W/W under greenhouse conditions. After soil amendment, agronomic growth parameters and soil parameters were analyzed. The degree of expression of phytoalexins and defense genes was carried out by UPLC-DAD- HRESI–MS (+) and qRT-PCR analyses, respectively. The results obtained show that soil treatment with these composites improves the sanitary quality of cocoa seedlings in controlled conditions through the stimulation of agronomic growth parameters such as plant height, number of leaves, leaf surface area, and fresh weight of shoot and root. The latter leads to an increase in soil pH by about 32 % as well as the improvement of the nutritional status of the soil by 90%. Moreover, bio-oyster shell soil amendment induces up-regulation of TcPer-1, TcGlu1, TcChiB, and TcMYBPA genes and accumulation of primed metabolites such as flavonols, brevifolin, and isoflavone compounds before and after infection. This stimulation based on biological substances make it possible to pre-immunize cocoa plants against pathogen attacks and would be a good candidate to improve the quality of cocoa seeds in Cameroon