ArticleAnalytical chemistry2017
Nanobody Based Immunoassay for Human Soluble Epoxide Hydrolase Detection Using Polymeric Horseradish Peroxidase (PolyHRP) for Signal Enhancement: The Rediscovery of PolyHRP?
Article in Analytical chemistry, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 30 papers.
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Who cites it
30 citing papers in PubMed, 67 citations in OpenAlex.
- Advances on Sensors and Diagnostics of SARS-CoV-2 Biomarkers.Analytical chemistry · 2026Article
- Advances in Immunological Methods for the Detection ofSensors (Basel, Switzerland) · 2026Review
- An immunoassay based on polyHRP signal amplification for HPV16/18 E6 RNA detection with anti-DNA-RNA hybrid antibody.Analytical sciences : the international journal of the Japan Society for Analytical Chemistry · 2025Article
- Development of Immunoassays for Foodborne Pathogenic Bacteria Detection Using PolyHRP for Signal Enhancement.Biosensors · 2025Article
- Nanobody based immunoassay for detection of aquatic virus: Giant salamander iridovirus.Analytica chimica acta · 2025Article
- Strategies for the Immobilization and Signal Amplification of a Double Nanobody Sandwich ELISA for Human Microsomal Epoxide Hydrolase.Analytical chemistry · 2024Article
- Article
- Modulating the sEH/EETs Axis Restrains Specialized Proresolving Mediator Impairment and Regulates T Cell Imbalance in Experimental Periodontitis.Journal of immunology (Baltimore, Md. : 1950) · 2024Article
- Nanobody based immunoassay for alpha fetal protein detection using streptavidin-conjugated polymerized horseradish peroxidase for signal amplification.Analytical sciences : the international journal of the Japan Society for Analytical Chemistry · 2023Article
- The Generation of a Nanobody-Based ELISA for Human Microsomal Epoxide Hydrolase.International journal of molecular sciences · 2023Article
- Effects of Individual Circulating FFAs on Plasma and Hepatic FFA Epoxides, Diols, and Epoxide-Diol Ratios as Indices of Soluble Epoxide Hydrolase Activity.International journal of molecular sciences · 2023Article
- Enhanced Competitive Immunomagnetic Beads Assay Assisted with PAMAM-Gold Nanoparticles Multi-Enzyme Probes for Detection of Deoxynivalenol.Biosensors · 2023Article
- PROTAC-Mediated Selective Degradation of Cytosolic Soluble Epoxide Hydrolase Enhances ER Stress Reduction.ACS chemical biology · 2023Article
- Mix-and-Read Nanobody-Based Sandwich Homogeneous Split-Luciferase Assay for the Rapid Detection of Human Soluble Epoxide Hydrolase.Analytical chemistry · 2023Article
- Molecular Targets for Foodborne Pathogenic Bacteria Detection.Pathogens (Basel, Switzerland) · 2023Review
- Structure and specificity of an anti-chloramphenicol single domain antibody for detection of amphenicol residues.Protein science : a publication of the Protein Society · 2022Article
- Soluble Epoxide Hydrolase Is Associated with Postprandial Anxiety Decrease in Healthy Adult Women.International journal of molecular sciences · 2022Article
- Improved ELISA for linoleate-derived diols in human plasma utilizing a polyHRP-based secondary tracer.Analytical methods : advancing methods and applications · 2022Article
- Horseradish peroxidase-triggered direct in situ fluorescent immunoassay platform for sensing cardiac troponin I and SARS-CoV-2 nucleocapsid protein in serum.Biosensors & bioelectronics · 2022Article
- A comprehensive comparison between camelid nanobodies and single chain variable fragments.Biomarker research · 2021Review
Corrections and comments
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Authors and funding
9 authors at 3 institutions in 2 countries.
Funding
Abstract
Soluble epoxide hydrolase (sEH) is a potential pharmacological target for treating hypertension, vascular inflammation, cancer, pain, and multiple cardiovascular related diseases. A variable domain of the heavy chain antibody (termed single domain antibody (sdAb), nanobody, or VHH) possesses the advantages of small size, high stability, ease of genetic manipulation, and ability for continuous manufacture, making such nanobody a superior choice as an immunoreagent. In this work, we developed an ultrasensitive nanobody based immunoassay for human sEH detection using polymeric horseradish peroxidase (PolyHRP) for signal enhancement. Llama nanobodies against human sEH were used as the detection antibody in sandwich enzyme linked immunosorbent assays (ELISA) with polyclonal anti-sEH as the capture antibody. A conventional sandwich ELISA using a horseradish peroxidase (HRP) labeled anti-hemeagglutinin (HA) tag as the tracer showed a marginal sensitivity (0.0015 optical density (OD)·mL/ng) and limit of detection (LOD) of 3.02 ng/mL. However, the introduction of the PolyHRP as the tracer demonstrated a 141-fold increase in the sensitivity (0.21 OD·mL/ng) and 57-fold decrease in LOD (0.05 ng/mL). Systematic comparison of three different tracers in four ELISA formats demonstrated the overwhelming advantage of PolyHRP as a label for nanobody based immunoassay. This enhanced sEH immunoassay was further evaluated in terms of selectivity against other epoxide hydrolases and detection of the target protein in human tissue homogenate samples. Comparison with an enzyme activity based assay and a Western blot for sEH detection reveals good correlation with the immunoassay. This work demonstrates increased competiveness of nanobodies for practical sEH protein detection utilizing PolyHRP. It is worthwhile to rediscover the promising potential of PolyHRP in nanobody and other affinity based methods after its low-profile existence for decades.
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Registered trials
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