Rapid, Visual Detection of Klebsiella pneumoniae Using Magnetic Nanoparticles and an Horseradish Peroxidase-Probe Based Immunosensor

Instrument-Free and Visual Detection of Salmonella Based on Magnetic Nanoparticles and an Antibody Probe Immunosensor

“…The titers of mAbs 2F1 and 1B12 were evaluated via ELISA, and the titers of both mAbs reached 1:204800 (Figure 2E,F). The K D values of mAb 2F1 and 1B12 were measured as described in our previous published study [22], and were calculated as K D = 3.677 ± 0.33 nM for mAb 2F1 and K D = 1.126 ± 0.15 nM for mAb 1B12.…”

“…The first one is that the surfaces of the NHS beads are coated with abundant mAb 2F1 that specifically recognize Salmonella (Figure 4A), which means the prepared INBs can efficiently and accurately enrich the Salmonella from different biological samples within 30 min, as shown in Figure 6A,B. Compared with the carboxylic modified magnetic beads used in our previous study and in other research articles [22], the NHS beads are coupled with antibodies with high efficiency and short timeframes, without needing EDC or glutaraldehyde for activation. The other advantage is that the mAb probe, having high affinity, can rapidly form a sandwich structure within 20 min, as shown in Figure 6C,D.…”

“…Conversely, immunodiagnostic approaches are more rapid, sensitive, and stable than nucleic acid assays, especially for samples such as milk [41], whole blood [42], and saliva [43]. Many novel immunosensors have been developed, which combine immunology with magnetic nanoparticles [22], platinum nanoparticles [44], or Pt nanomotors [45]. Instrument-free and mobile diagnostic technologies could transform the current foodborne pathogen detection systems, particularly in resource-limited settings.…”

“…The HRP mAb probes were synthesized using mAbs coupled with HRP. All probes were prepared as described in our previous study [22]. Briefly, 10 mg of HRP was dissolved in 1 mL of 0.1 M NaHCO 3 and then oxidized for 2 h with 1 mL of 10 mM NaIO 4 .…”

Visual and Rapid Detection of Klebsiella pneumoniae by Magnetic Immunocapture-Loop-Mediated Isothermal Amplification Assay

“…Then, three mAbs were generated and designated as 2F1, 1E6, and 4E6, after cell fusion and subcloning. The titer, subtypes, and K D value of 2F1, 1E6, and 4E6 were determined in our previous study (26). In the next step, mAb 1E6, which had the highest affinity, was purified and confirmed by SDS-PAGE ( Figure 2B).…”