![]() The team also generated a point mutant in the S-BAT binding module at Asparagine30 to form S-BAT*. The molecule representing the best linker combination was named S-BAT. The researchers plotted the performance obtained based on the detection of one nano-molar (nM) recombinant S in phosphate-buffered saline (PBS). The signal to noise (S/N) ratio multiplied by the signal change (S-N) indicated the performance for each linker composition. Its N- and C-termini are set ~25 angstroms (Å) apart and in opposite directions, thus making a rigid scaffold for minimizing the effective concentration of the fused split luciferase halves. In addition, LCB1 is rigid, thermally stable, and has no disulfide bonds to confound purification. LCB1 binds to the receptor-binding domain (RBD) of SARS-CoV-2 S protein with a dissociation constant (K d) of 10-10. The researchers selected the computationally designed long-chain base subunit 1(LCB1) of serine palmitoyltransferase as the binding module. BATs consist of a binding module for antigen and a split enzyme, NanoBiT, that fuse in tandem to the N and C terminus of the binding module. In the present study, researchers deployed a single-component, luminescent BAT biosensor for detecting the SARS-CoV-2 S protein in multiple assay formats. Compared to other single component platforms, they do not count on large conformational changes in the binding module or competition with a tethered decoy. Moreover, they only need a camera for detection and are adapted to multiple assay formats. Split luciferase enzyme-based single-component luminescent biosensors are simpler to produce, highly sensitive, and easier to use. ![]() Iii) they require a minimum of two unique antibodies to function. Ii) most antibody-based assays require chemical modification to immobilize the antibodies, and I) antibodies need mammalian expression systems for production, ![]() These technologies have three limitations, as follows: Image Credit: NIAID BackgroundĪll the discovery and translational biomedical research depend primarily on antibody-based technologies for small-molecule and protein analyte detection. Study: A single-component luminescent biosensor for the SARS-CoV-2 spike protein. In a recent study posted to the bioRxiv* pre-print server, researchers developed a novel Binding Activated Tandem split enzyme (BAT) biosensor for the detection of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike (S) protein. ![]() By Neha Mathur Reviewed by Danielle Ellis, B.Sc. ![]()
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