Rapid and efficient isolation of intact lysosomes is necessary to study their functions and metabolites by proteomic analysis. We developed a swift and robust nanoparticle-based magnetic separation method in which magnetic− plasmonic hybrid nanoparticles (MPNPs) conjugated with amino dextran (aDxt) were targeted to the lumen of lysosomes via the endocytosis pathway. For well-directed magnetic separation of the lysosomes, it is important to trace the intracellular trafficking of the aDxt-conjugated MPNPs (aDxt-MPNPs) in the endocytosis pathway. Therefore, we analyzed the intracellular transport process of the aDxt-MPNPs by investigating the time-dependent colocalization of plasmonic scattering of aDxt-MPNPs and immunostained marker proteins of organelles using the threshold Manders' colocalization coefficient (R t ). Detailed analysis of time variations of R t for early and late endosomes and lysosomes allowed us to derive the transport kinetics of aDxt-MPNPs in a cell. After confirming the incubation time required for sufficient accumulation of aDxt-MPNPs in lysosomes, the lysosomes were magnetically isolated as intact as possible. By varying the elapsed time from homogenization to complete isolation of lysosomes (t delay ) and temperature (T), the influences of t delay and T on the protein composition of the lysosomes were investigated by polyacrylamide gel electrophoresis and amino acid analysis. We found that the intactness of lysosomes could become impaired quite quickly, and to isolate lysosomes as intact as possible with high purity, t delay = 30 min and T = 4 °C were optimal settings.

For lateral flow immunoassay (LFIA), it is an important challenge to enhance the detection sensitivity to the same level as polymerase chain reaction or enzyme-linked immunosorbent assay to make LFIA pervasive in the field of on-site environmental analysis. We recently demonstrated that the LFIA sensitivity is dramatically enhanced by using Pt-nanoparticle−latex nanocomposite beads (Pt-P2VPs) as probes for the detection of the influenza A (H1N1) antigen compared with using conventional Au colloids as probes. Here, to further enhance the LFIA sensitivity using Pt-P2VPs, superparamagnetic iron oxide nanoparticles (SPIONs) were chemically conjugated to Pt-P2VPs (Pt-P2VP@SPION) to give them magnetic separation capability (enrichment and/or purification). To investigate the effect of magnetic enrichment on the LFIA sensitivity in a sandwich format, the C-reactive protein (CRP) was chosen as a model analyte and anti-CRP antibody (CRPAb)-conjugated Pt-P2VP@SPION (Pt-P2VP@SPION-CRPAb) beads were used as probes. The visual limit of detection (LOD) of LFIA was successfully lowered by increasing the magnetic enrichment factor φ. The minimum LOD under the present experimental conditions was 0.08 ng/mL for φ = 40, which is 26-fold lower than that of the standard Au-nanoparticle-based LFIA. In theory, the LOD can be unlimitedly decreased by just increasing φ. However, the times required for both the antigen−antibody binding reaction and magnetic separation dramatically increase with φ. We also propose solutions to overcome this drawback.

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Designing high-performance TiO2-based photocatalysts with reduced bandgap is obligatory towards utilizing abundant sunlight. In this study, a novel hybrid was fabricated by interfacial polymerization of polyaniline (PANi) and polypyrrole (PPy)...

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