As substitutes for natural peroxidases, most nanomaterial-based enzyme mimetics (nanozymes) have unique properties such as high stability, low-cost, large surface area, and high catalytic activity. However, they usually work in acidic conditions and thus impede their real applications. In this work, by modulating the nanostructure, composition, and surface property of the bimetallic materials, the positively charged poly(diallyldimethylammonium)-stabilized Au@Ag heterogeneous nanorods (NRs) were developed as synergistic peroxidase-like interfaces, which exhibited high activity over a wide pH range (pH 4.0-6.5) using 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) diammonium salt (ABTS) as the chromogenic substrate. At pH 6.5, the peroxidase-like activity for the Au@Ag heterogeneous NRs was stable and optimal within 20-40 °C. Moreover, the Au@Ag heterogeneous NRs showed excellent temperature stability and long-term storage stability. Given these characters, the detection of H2O2 at pH 6.5 was proposed on the basis of the Au@Ag heterogeneous NRs catalyzing the colorimetric reaction of H2O2 and ABTS, where the oxidized ABTS showed a typical absorption peak at 414 nm. The absorbance at 414 nm was linear with H2O2 concentration from 0.01 to 10 mM. Further, considering that Au@Ag heterogeneous NRs and glucose oxidase (GOx) have similar optimal pH for catalytic activities, a novel one-pot method for the detection of glucose was developed by the coupled catalytic reaction using GOx, Au@Ag heterogeneous NRs, and ABTS at nearly neutral pH (pH 6.5) and 37 °C. This proposed method had simple and rapid processes, wide linear range (0.05-20 mM), and reliability for the successful analysis of real samples. On the basis of these attractive and unique characteristics, Au@Ag heterogeneous NRs can become promising substitutes for peroxidase in analytical chemistry and environmental science.

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Probes against targets can be selected from the landscape phage library f8/8, displaying random octapeptides on the pVIII coat protein of the phage fd-tet and demonstrating many excellent features including multivalency, stability, and high structural homogeneity. Prostate-specific antigen (PSA) is usually determined by immunoassay, by which antibodies are frequently used as the specific probes. Herein we found that more advanced probes against free prostate-specific antigen (f-PSA) can be screened from the landscape phage library. Four phage monoclones were selected and identified by the specificity array. One phage clone displaying the fusion peptide ERNSVSPS showed good specificity and affinity to f-PSA and was used as a PSA capture probe in a sandwich enzyme-linked immunosorbent assay (ELISA) array. An anti-human PSA monoclonal antibody (anti-PSA mAb) was used to recognize the captured antigen, followed by horseradish peroxidase-conjugated antibody (HRP-IgG) and o-phenylenediamine, which were successively added to develop plate color. The ELISA conditions such as effect of blocking agent, coating buffer pH, phage concentration, antigen incubation time, and anti-PSA mAb dilution for phage ELISA were optimized. On the basis of the optimal phage ELISA conditions, the absorbance taken at 492 nm on a microplate reader was linear with f-PSA concentration within 0.825-165 ng/mL with a low limit of detection of 0.16 ng/mL. Thus, the landscape phage is an attractive biomolecular probe in bioanalysis.