Optimized immobilization of single chain variable fragment antibody onto non-toxic fluorescent nanoparticles for efficient preparation of a bioprobe

Abstract

Single-chain variable fragment antibody (scFv) is a small molecular weight antibody that can be used for both therapeutic and diagnostic purposes. To visualize the interaction with the target biomolecules, scFv must be labeled with fluorescent molecules. In this study, to achieve the efficient labeling of scFv, we developed scFv-fluorescent nanoparticle conjugates to utilize scFv as bioprobes. As fluorescent carriers, cadmium-free ZnS-AgInS₂/ZnS core/shell nanoparticles were used, and scFv was immobilized onto the nanoparticles via the interaction of nickel ions on nitrilotriacetic acid and hexahistidine (His-tag) fused with scFv. UV-Vis, fluorescence spectra, NMR, and dynamic laser scattering were used to characterize the scFv immobilized fluorescent nanoparticles (scFv-FNPs). The amounts of scFv on FNPs were controlled by the concentration of scFv. The scFv-FNPs that were prepared were non-toxic and selectively bound to cancer cells. The scFv-FNPs could be used as bioanalytical tools, and the immobilization method described here is a promising method for labeling biomolecules with the His-tag.

Keywords: Antibody; Fluorescent probe; Nanoparticles; Quantum dots; ScFv.