Abstract
Soybean peroxidase (SBP) is characterized by relatively high heat resistance, wide substrate scope, good stability and broad pH applicability. These advantages led SBP to be used as labeledantibody for immunoassays. In this work, monoclonal antibody was linked with SBP to beta-HCG (Mab II-SBP) for enzyme-linked immunosorbent assay (ELISA) or chemiluminescence assay to explore its potential as an alternative to the widely used horseradish peroxidase (HRP) based ELISA. The Mab II-SBP was prepared by the sodium periodate oxidation method. The effects of enzyme coupling rate and molar ratio of Mab II to enzyme on assay performance were investigated. The optimum mass concentration ratio of Mab II and SBP was determined to be 1:3. The best coat concentration of the monoclonal antibody to alfa-HCG antibody (Mab I) was 4 μg/mL. Under optimum conditions, the HCG concentration was determined by the proposed ELISA assay with linear response range from 0.18 to 18 ng/mL and detection limit of 0.07 ng/mL at a signal-to-noise ratio of 3. Both enhancers, 3-(10’-phenothiazinyl) propane- 1-sulfonate (SPTZ) and 4-morpholinopyridine (MORPH), amplified chemiluminescence intensity nearly 200-fold in comparison to the SBP-luminol-H2O2 system. The present work demonstrates the feasibility and rationale of SBP-labeling antibodies for ELISA and chemiluminescence assays.
Keywords: Chemiluminescence, ELISA, enhancers, enzyme-labeled antibody, immunoassay, soybean peroxidase.
Current Analytical Chemistry
Title:The Enzyme Linked Immunosorbent and Chemiluminescence Assay for the Detection of Human Chorionic Gonadotrophin Using Soybean Peroxidase as Label Enzyme
Volume: 11 Issue: 2
Author(s): Fang Zhao, Wei Zhu, Juan Su, Yanchun Xue, Wei Wei and Songqin Liu
Affiliation:
Keywords: Chemiluminescence, ELISA, enhancers, enzyme-labeled antibody, immunoassay, soybean peroxidase.
Abstract: Soybean peroxidase (SBP) is characterized by relatively high heat resistance, wide substrate scope, good stability and broad pH applicability. These advantages led SBP to be used as labeledantibody for immunoassays. In this work, monoclonal antibody was linked with SBP to beta-HCG (Mab II-SBP) for enzyme-linked immunosorbent assay (ELISA) or chemiluminescence assay to explore its potential as an alternative to the widely used horseradish peroxidase (HRP) based ELISA. The Mab II-SBP was prepared by the sodium periodate oxidation method. The effects of enzyme coupling rate and molar ratio of Mab II to enzyme on assay performance were investigated. The optimum mass concentration ratio of Mab II and SBP was determined to be 1:3. The best coat concentration of the monoclonal antibody to alfa-HCG antibody (Mab I) was 4 μg/mL. Under optimum conditions, the HCG concentration was determined by the proposed ELISA assay with linear response range from 0.18 to 18 ng/mL and detection limit of 0.07 ng/mL at a signal-to-noise ratio of 3. Both enhancers, 3-(10’-phenothiazinyl) propane- 1-sulfonate (SPTZ) and 4-morpholinopyridine (MORPH), amplified chemiluminescence intensity nearly 200-fold in comparison to the SBP-luminol-H2O2 system. The present work demonstrates the feasibility and rationale of SBP-labeling antibodies for ELISA and chemiluminescence assays.
Export Options
About this article
Cite this article as:
Zhao Fang, Zhu Wei, Su Juan, Xue Yanchun, Wei Wei and Liu Songqin, The Enzyme Linked Immunosorbent and Chemiluminescence Assay for the Detection of Human Chorionic Gonadotrophin Using Soybean Peroxidase as Label Enzyme, Current Analytical Chemistry 2015; 11 (2) . https://dx.doi.org/10.2174/1573411011666141223220346
DOI https://dx.doi.org/10.2174/1573411011666141223220346 |
Print ISSN 1573-4110 |
Publisher Name Bentham Science Publisher |
Online ISSN 1875-6727 |
Call for Papers in Thematic Issues
Advancements in biomedicine, corrosion inhibition/monitoring, and lubrication of carbon dots
Carbon dots, a class of carbon-based nanomaterials, have garnered substantial interest in various scientific and engineering fields due to their unique properties, including excellent optical properties, outstanding biocompatibility, easy preparation, good dispersity, ultrasmall size, low toxicity and environmental friendliness. This special issue of Current Analytical Chemistry aims to showcase cutting-edge ...read more
Analytical methods for environmental and food analysis
In recent years, the study of the chemical composition of food and the environment depends on analytical methods in order to acquire information about the chemical composition, processing, quality control and contamination of soil, water, air, plants and foods, aspects very important for food safety and environmental quality control. There ...read more
Applications of artificial intelligence in chemical analysis
Artificial intelligence (AI) has demonstrated its versatility in almost all scientific fields, especially analytical chemistry, which is leading the way in the integration of AI. According to Chemical Abstract Services, one of the major disciplines contributing to AI-related publications and patents is analytical chemistry. That?s because techniques such as deep ...read more
Bioanalytical and Biosensor Technologies for Target Molecules
Bioanalytical and biosensor technologies constitute the core of detecting and sensing diverse molecules within living organisms, encompassing proteins, DNA, small molecular metabolites, and beyond. These molecules assume pivotal roles in biological processes, rendering their analysis and sensing indispensable for comprehending biological phenomena and unraveling disease mechanisms. In the current landscape ...read more
- Author Guidelines
- Graphical Abstracts
- Fabricating and Stating False Information
- Research Misconduct
- Post Publication Discussions and Corrections
- Publishing Ethics and Rectitude
- Increase Visibility of Your Article
- Archiving Policies
- Peer Review Workflow
- Order Your Article Before Print
- Promote Your Article
- Manuscript Transfer Facility
- Editorial Policies
- Allegations from Whistleblowers
Related Articles
-
Tubulins as Therapeutic Targets in Cancer: from Bench to Bedside
Current Pharmaceutical Design Electrochemical Examination of Nanomaterial Modified Carbon Based Electrodes
Current Analytical Chemistry Stress, Aging and Reliability of Antioxidant Enzyme Defense
Current Aging Science Current and Emerging Systemic Therapy in Gastro-Esophageal Cancer “The Old and New Therapy for Metastatic Disease, The Role of Adjuvant and Neoadjuvant Therapy for Localized Disease”
Current Clinical Pharmacology Chronic Diseases and COVID-19: A Review
Endocrine, Metabolic & Immune Disorders - Drug Targets Using Protein-protein Interaction Network Information to Predict the Subcellular Locations of Proteins in Budding Yeast
Protein & Peptide Letters Mannosylated Solid Lipid Nanocarriers of Chrysin to Target Gastric Cancer: Optimization and Cell Line Study
Current Drug Delivery Quantification of Several Bioactive Compounds and Antioxidant Activities of Six Cultivars of Brazilian Blueberry
The Natural Products Journal Targeting the Chemokine Receptor CXCR3 and Its Ligand CXCL10 in the Central Nervous System: Potential Therapy for Inflammatory Demyelinating Disease?
Current Neurovascular Research Structure and Mechanism of Action of Tau Aggregation Inhibitors
Current Alzheimer Research The Use of Keratin in Biomedical Applications
Current Drug Targets Aryl- and Heteroaryl-Thiosemicarbazone Derivatives and Their Metal Complexes: A Pharmacological Template
Recent Patents on Anti-Cancer Drug Discovery Apoptosis Signalling Activated by TNF in the Lower Gastrointestinal Tract-Review
Current Pharmaceutical Biotechnology Strategies for the Assessment of Metabolic Profiles of Steroid Hormones in View of Diagnostics and Drug Monitoring: Analytical Problems and Challenges
Current Drug Metabolism Allosteric Modulation of Nicotinic Acetylcholine Receptors: The Concept and Therapeutic Trends
Current Pharmaceutical Design Discrepant mRNA and Protein Expression in Immune Cells
Current Genomics High-Throughput Screening Assays to Discover Small-Molecule Inhibitors of Protein Interactions
Current Drug Discovery Technologies Carbohydrate Mimotopes in the Rational Design of Cancer Vaccines
Current Topics in Medicinal Chemistry Nanomedicine: Magnetic Nanoparticles and their Biomedical Applications
Current Medicinal Chemistry Can Radiology Replace Endoscopy in IBD?
Current Drug Targets