Biomimetic -mineralized multifunctional nanoflowers for anodic-stripping voltammetric immunoassay of rehabilitation-related proteins.

C-reactive proteins (CRPs; an acute-phase protein) in patients with initial acute cerebral infarction neurological rehabilitation prediction have a significant correlation. In this work, a simple and sensitive anodic-stripping voltammetric (ASV) immunosensing system was innovatively designed for the quantitative screening of target CRPs using biomimetic-mineralized bifunctional antibody-Cu3(PO4)2 nanoflowers as molecular tags. In this system, a monoclonal anti-CRP antibody-anchored microtiter plate was utilized to specifically capture target CRPs from the sample. For detection, a sandwiched immunoreaction mode was employed with the antibody-Cu3(PO4)2 nanoflowers in the presence of analytes. Subsequent ASV measurement of copper ions (Cu2+) released under acidic conditions from the bifunctional nanoflowers was conducted at an in situ prepared mercury film electrode. The introduction of hybrid nanoflowers greatly increased the loading amount of copper ions on the molecular tag, thereby amplifying the detectable signal of electrochemical immunoassay. Meanwhile, factors influencing the analytical properties of the electrochemical immunoassay were investigated in detail. By combining the high-efficiency nanohybrids with signal amplification, the dynamic concentration range of electrochemical immunoassay spanned from 0.01 ng mL-1 to 100 ng mL-1 toward the target CRP. The limit of detection was calculated to be 0.0079 ng mL-1 at 3Sblank criterion. Intra- and interassay imprecisions (relative standard deviations: RSDs) were less than or equal to 6.72%. Good anti-interference ability, long-term storage stability, and acceptable accuracy for the evaluation of human serum specimens were observed during a series of procedures to determine the target protein. In addition, the bifunctional nanoflower-based immunosensing system offers promise for the simple, cost-effective analysis of disease-related proteins.