Development of an elisa for the diagnosis of reactive IgE antibodies anti-therapeutic horse sera.
Abstract: Hypersensitive diseases that involve IgE reactivity are important concern of public, especially those encompassing the potential pathogenesis from the administration of horse serum-based therapeutics such as antivenoms. A method for the definitive diagnosis of reactive IgE is important for identifying allergic patients to control severe collateral effects during planned and emergency application of immunotherapies when the allergy source cannot be avoided for treatment. To date, no tests have been developed to accompany the wide range of antivenoms produced from horse sera. The aim of this was to develop a cost-effective ELISA of high sensitivity and specificity to detect circulating patient IgE that binds horse IgG, the most prevalent antibody class in passive antibody therapies. Horse IgG was purified in a single step on jacalin-Sepharose and absorbed to standard ELISA plates as the capture molecule for reactive human IgE. The direct performance evaluation with allergenic and non-allergenic patient, together with competitive peptides assays, showed high sensitivity and specificity to detect human IgE that recognized horse IgG. The analytical sensitivity and ED were calculated to be 0.01 μg mL and 0.052 μg mL, respectively. The intra- and inter-assay coefficient of variation ranged from 3.3 to 11.1% and 4.0-8.0%, respectively. The horse IgG-based ELISA assay can detect reactive allergenic IgE at picomolar concentrations. The coefficient of variation suggests that it can be easily standardized between laboratories, provide rapid and can be applied to population surveillance. Patient management during treatment for envenomation would be greatly improved by a robust and reliable diagnostic test for preexisting allergies to mitigate life-threating consequences of hypersensitivity.
Copyright © 2017 Elsevier Ltd. All rights reserved.
Publication Date: 2017-08-12 PubMed ID: 28811248DOI: 10.1016/j.toxicon.2017.08.012Google Scholar: Lookup
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- Journal Article
Summary
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The research article outlines the development of a highly sensitive and specific ELISA (Enzyme-Linked Immunosorbent Assay) to detect reactive IgE (Immunoglobulin E) antibodies against therapeutic horse sera in patients, which is crucial in managing allergic reactions during treatments that cannot avoid the use of horse serum-based therapies like antivenoms.
Objective of the Research
- The study aimed to design an affordable ELISA with high sensitivity and specificity to detect reactive IgE antibodies against horse IgG (Immunoglobulin G), a major antibody type utilized in passive antibody therapies.
Methodology
- Horse IgG was initially purified in a single step on jacalin-Sepharose (a carbohydrate-binding protein) and then adsorbed to standard ELISA plates to serve as the capture molecule for reactive human IgE.
- The performance of this newly developed assay was evaluated using serum samples from both allergenic and non-allergenic patients, along with competitive peptides assays.
Findings of the Study
- The assay showed high sensitivity and specificity in detecting human IgE that recognized horse IgG, indicating its potential usage in identifying patients allergic to therapeutic horse sera.
- The test could identify reactive allergenic IgE at picomolar concentrations, demonstrating its high sensitivity.
- The intra- and inter-assay coefficient of variation ranged from 3.3 to 11.1% and 4.0 to 8.0% respectively, suggesting that the assay can be easily standardized across different labs.
Significance of the Research
- This research is critical as no tests have been created so far to go along with the wide array of antivenoms derived from horse sera.
- The ability to rapidly identify patients who are allergic to horse serum-based therapies can potentially prevent severe side effects or hypersensitive reactions during the application of these therapies.
- Management of envenomation treatment can be significantly improved and life-threatening consequences could be mitigated with a reliable diagnostic test for preexisting allergies.
Cite This Article
APA
De-Simone SG, Souza ALA, Aguiar AS, Melgarejo AR, Provance DW.
(2017).
Development of an elisa for the diagnosis of reactive IgE antibodies anti-therapeutic horse sera.
Toxicon, 138, 37-42.
https://doi.org/10.1016/j.toxicon.2017.08.012 Publication
Researcher Affiliations
- FIOCRUZ, Center of Technological Development in Health (CDTS)/National Institute of Science and Technology for Innovation on Neglected Diseases (INCT-IDN), Rio of Janeiro, RJ, Brazil; FIOCRUZ, Oswaldo Cruz Institute, Laboratory of Experimental and Computational Biochemistry of Pharmaceuticals, Rio de Janeiro, RJ, Brazil; Federal Fluminense University, Biology Institute, Department of Cellular and Molecular Biology, Niterói, Rio Janeiro, Brazil. Electronic address: dsimone@cdts.fiocruz.br.
- FIOCRUZ, Oswaldo Cruz Institute, Laboratory of Experimental and Computational Biochemistry of Pharmaceuticals, Rio de Janeiro, RJ, Brazil.
- Vital Brazil Institute, Research and Development Laboratory, Niterói, RJ, Brazil.
- Vital Brazil Institute, Research and Development Laboratory, Niterói, RJ, Brazil.
- FIOCRUZ, Center of Technological Development in Health (CDTS)/National Institute of Science and Technology for Innovation on Neglected Diseases (INCT-IDN), Rio of Janeiro, RJ, Brazil.
MeSH Terms
- Animals
- Antibodies, Anti-Idiotypic / analysis
- Antivenins / immunology
- Enzyme-Linked Immunosorbent Assay / methods
- Horses
- Humans
- Hypersensitivity / diagnosis
- Immunoglobulin G / analysis
- Immunoglobulin G / immunology
- Sensitivity and Specificity
Citations
This article has been cited 6 times.- De-Simone SG, Napoleão-Pêgo P, Lechuga GC, Carvalho JPRS, Gomes LR, Cardozo SV, Morel CM, Provance DW Jr, Silva FRD. High-Throughput IgG Epitope Mapping of Tetanus Neurotoxin: Implications for Immunotherapy and Vaccine Design.. Toxins (Basel) 2023 Mar 24;15(4).
- De-Simone SG, Napoleão-Pêgo P, Gonçalves PS, Lechuga GC, Cardoso SV, Provance DW Jr, Morel CM, da Silva FR. B-Cell Epitope Mapping of the Vibrio cholera Toxins A, B, and P and an ELISA Assay.. Int J Mol Sci 2022 Dec 28;24(1).
- De-Simone SG, Napoleão-Pêgo P, Gonçalves PS, Lechuga GC, Mandonado A Jr, Graeff-Teixeira C, Provance DW Jr. Angiostrongylus cantonensis an Atypical Presenilin: Epitope Mapping, Characterization, and Development of an ELISA Peptide Assay for Specific Diagnostic of Angiostrongyliasis.. Membranes (Basel) 2022 Jan 19;12(2).
- Napoleão-Pêgo P, Carneiro FRG, Durans AM, Gomes LR, Morel CM, Provance DW Jr, De-Simone SG. Performance assessment of a multi-epitope chimeric antigen for the serological diagnosis of acute Mayaro fever.. Sci Rep 2021 Jul 28;11(1):15374.
- De-Simone SG, Gomes LR, Napoleão-Pêgo P, Lechuga GC, de Pina JS, da Silva FR. Epitope Mapping of the Diphtheria Toxin and Development of an ELISA-Specific Diagnostic Assay.. Vaccines (Basel) 2021 Mar 26;9(4).
- Aimulajiang K, Naqvi MA, Chu W, Lu M, Tian X, Bu Y, Memon MA, Li X, Xu L, Song X, Yan R. Adhesion-Regulating Molecule from Haemonchus contortus: Potential Antigen for Diagnosis of Early Infection in Goats.. Pathogens 2019 Dec 30;9(1).
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