A Genetically Engineered Biofilm Material for SARS‐CoV‐2 Capturing and Isolation

Abstract

The novel severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) is continuously infecting people all around the world since its outbreak in 2019. Studies for numerous infection detection strategies are continuing. The sensitivity of detection methods is crucial to separate people with mild infections from people who are asymptomatic. In this sense, a strategy that would help to capture and isolate the SARS‐CoV‐2 virus prior to tests can be effective and beneficial. To this extent, genetically engineered biomaterials grounding from the biofilm protein of Escherichia coli are beneficial due to their robustness and adaptability to various application areas. Through functionalizing the E. coli biofilm protein, diverse properties can be attained such as enzyme display, nanoparticle production, and medical implant structures. Here, E. coli species are employed to express major curli protein CsgA and Griffithsin (GRFT) as fusion proteins, through a complex formation using SpyTag and SpyCatcher domains. In this study, a complex system with a CsgA scaffold harboring the affinity of GRFT against Spike protein to capture and isolate SARS‐CoV‐2 virus is successfully developed. It is shown that the hybrid recombinant protein can dramatically increase the sensitivity of currently available lateral flow assays for Sars‐CoV‐2 diagnostics.

Rational design of functional amyloid fibrillar assemblies

“…This is achieved through the interaction between the functional curli fibrils and the spike proteins of the targeted viruses. 93,165,166 The recombinant biofilms could effectively concentrate and detect viruses in waste water with higher accuracy compared to traditional methods. By growing the engineered biofilms on commercial polypropylene filters, the modified filters held great potential as a convenient device for waste water treatment.…”

An Antibiotic Degrading Engineered Living Material Platform to Combat Environmental Antibiotic Resistance

“…The preparation of CsgA-SpyTag fibers were conducted as described in our previous paper [21]. Briefly, ΔcsgA cells containing pZa tetO CsgA-SpyTag plasmid were inoculated into lysogeny broth (LB) medium supplied with ampicillin and incubated overnight at 37 °C, 200 rpm.…”

“…The SpyTag-SpyCatcher complex formation using the CsgA-SpyTag and SpyCatcher-CotA, SpyCatcher-YlmD subunits was conducted as described earlier in our previous work and also by Joshi et al [21,34]. Briefly, CsgA-SpyTag fibers were weighed and aliquoted to have 1 mg/mL final concentration following the induction and filtration processes explained previously.…”

“…The CsgA-SpyTag fibers were confirmed using Transmission Electron Microscopy (TEM) (Figure 2). The CsgA-SpyTag biofilm fibers were analyzed using the same methods in our previous work [21].…”

“…The approach for forming complex structures using SpyTag and SpyCatcher subunits is achieved using the same mechanism as explained in our previous work [21]. Briefly, an irreversible isopeptide bond formation is promoted in a favorable reaction medium between the unprotonated amine group of Lys 31 and carbonyl of Asp 117 [24].…”