A solid phase NMR-based sandwich immunoassay for the prostate-specific antigen (PSA) is presented. Carbon-encapsulated iron nanoparticles were functionalized with bovine serum albumin, coupled to monoclonal antibodies, and then used as magnetic labels. A nitrocellulose membrane with 8-μm pores was coated with capture antibodies and subsequently incubated with a serum sample and a suspension of the nanoconjugate. Test strips were placed in a portable homemade NMR relaxometer. Magnetic nanoparticles attached to nitrocellulose decrease the T2 relaxation time of the water protons located inside the pores of the membrane. Thus, T2 is inversely proportional to the concentration of the antigen (PSA) in the sample. The assay can be performed within 4 h. The detection limit is 0.44 ng mL-1. Kallikrein 2, human chorionic gonadotropin, and α-fetoprotein do not interfere. Graphical abstractSchematic representation of NMR relaxometry-based sandwich dot blot immunoassay of a prostate-specific antigen (PSA). Magnetic nanoparticles bound to immunosorbent decrease the transverse relaxation times (T2) of the water protons located within the pores of the membrane. RF coil: radiofrequency coil.
Keywords: Dot blot; Dot-immunoassay; Nitrocellulose; Relaxivity; Relaxometry.