Nonstructural protein 1 (NS1) functions in the replication of positive-sense RNA genomes of flaviviruses, such as Zika virus, and is highly homologous among flaviviruses. Glycosylated NS1 can be found in the blood of infected individuals up to nearly two weeks after infection, therefore being a potential target for flavivirus diagnosis. One option for detection of NS1 in the bloodstream could entail taking advantage of the carbohydrate-binding properties of lectins. Lectins have been isolated from a variety of eukaryotes and prokaryotes and often bind to carbohydrate chains with high specificity. The objective of this project is to determine the binding of several recombinantly-produced lectins to various flavivirus NS1 proteins and subsequently design biosensors for each NS1 using the optimal lectin as the recognition element. Lectins have been cloned using standard molecular biology techniques, expressed, and purified. The binding affinity and selectivity of lectins to each flavivirus NS1 were determined using Enzyme-linked immunosorbent assay (ELISA) and Surface Plasmon Resonance (SPR). Preliminary results suggest Avaren-Fc, an actinomycete-derived lectin actinohivin mutant dimerized by genetic fusion to the Fc region of human IgG1, and Zika NS1 may be an optimal lectin-NS1 pair. Avaren-Fc binds with less affinity to NS1 from other flaviviruses (dengue and yellow fever) and has not been found to bind to Japanese encephalitis virus NS1. Further work includes identifying other lectin-NS1 pairs, characterizing this binding by ELISA and SPR, and determining the feasibility of using lectins for detection of NS1 in bodily fluids by utilizing spiked samples of serum, urine, and saliva. In conclusion, employing optimal lectin-NS1 pairs, a biosensor will be designed as a rapid screening tool for flaviviruses. Many communities afflicted by flaviviruses could benefit from the introduction of a more rapid and cost-effective diagnostic tool with such high specificity as predicted from lectins.

Development of lectin-based biosensors for the detection of flaviviruses

2019 Spring National Meeting

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