Hemoglobin as a Capture Agent for Lipopolysaccharide Antigens in the Enzyme Immunoassay of Gram Negative Bacteria

Abstract

The affinity of hemoglobin for lipopolysaccharides (LPS) was exploited in its use as an inexpensive capture agent for LPS antigens in the enzyme immunoassay (EIA) of Gram negative bacteria. Two EIA formats were examined. In one, the macroporous solid phase PolymacronTM coated with hemoglobin was used to capture cholate‐heat extracted LPS antigens from broth cultures of representative Gram negative bacteria, including different Salmonella serotypes, which were then detected immunoenzymatically using specific antibodies. This provided a rapid, simple and inexpensive dot blot assay for these bacteria which minimized the requirement for laboratory equipment. In another format, a microtiter plate EIA was developed in which cholate‐heat extracted Salmonella. LPS antigens were captured in hemoglobin‐coated wells. The microtiter plate format is automatable and will therefore be useful in laboratories with high sample throughputs. While most of the results reported here focus on the detection of Salmonella spp., we also demonstrate the applicability of this system in the assay of Escherichia coli O157 LPS antigens.

Multifunctional Magnetic Copper Ferrite Nanoparticles as Fenton-like Reaction and Near-Infrared Photothermal Agents for Synergetic Antibacterial Therapy

“…The phenomenon reflected that magnetic enrichment significantly enhanced the photothermal effect of Hb-CFNPs, thereby reducing the concentration of Hb-CFNPs served as a photothermal agent and improving the biosafety. Additionally, it was worth mentioning that Hb-CFNPs could capture bacteria via the electrostatic force and the affinity of Hb for lipopolysaccharides of a bacterial cell wall. , Under magnetic mediation, bacteria was enriched (Figure S3), resulting in a significant increase in photothermal sterilization efficiency. Thus, Hb-CFNPs could be exploited as an ideal photothermal agent for ablation of bacteria.…”

“…lipopolysaccharides of a bacterial cell wall. 28,38 Under magnetic mediation, bacteria was enriched (Figure S3), resulting in a significant increase in photothermal sterilization efficiency. Thus, Hb-CFNPs could be exploited as an ideal photothermal agent for ablation of bacteria.…”

Development of aptamer-based capture and detection systems for the identification of analytes

“…(Blais & Yam azaki, 1991). The use o f hem oglobin as a capture agent for LPS has also been em ployed for the detection o f Salmonella in food sam ples (Blais et al, 1998). However,…”