Construction of an enzyme-based all-fiber SPR biosensor for detection of enantiomers

Chiral analysis of amino acids (AAs) is of great importance in medical science due to the distinctive effect of AA isomers on human health. Although various chiral recognition techniques have been developed, the quantitative chiral recognition of low-level AA isomers remains challenging. Here, we combined the fiber optic SPR with an enzyme-substrate recognition mechanism to construct a direct-assay-type chiral AA biosensor. As a proof-of-concept attempt, a recently discovered Rasamsonia emersonii D-amino acid oxidase (ReDAAO) with a wide substrate spectrum and high stability was immobilized on the graphene oxide and gold nanorods composites (GO-AuNRs), using both EDC/NHS coupling and the gold-binding peptide (GBP) method. Such a biosensor can distinguish two AA isomers at the same concentration. It achieved specific detection of D-amino acids (D-AAs) with a linear range from 5x10-4 mM to 30 mM. Furthermore, it showed good resistance to enantiomeric interference. When the percentage of D-AA increases in the isomer mixture, a good linear relationship between the D/(D + L)-AA ratio and SPR spectral shift was obtained. This unique combination of the enzyme, nanocomposite, and SPR taps into the rich reservoir of proteins for chiral receptors. It lays the foundation for protein-based chiral recognition of other clinically important small molecules in future biosensor designs.