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Identification of a common Ara h 3 epitope recognized by both the capture and the detection monoclonal antibodies in an ELISA detection kit.

Title: Identification of a common Ara h 3 epitope recognized by both the capture and the detection monoclonal antibodies in an ELISA detection kit.
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Name(s): Zhao, Lipei, author
Zhao, Liang, author
Zhang, Buchang, author
Robotham, Jason M, author
Roux, Kenneth H, author
Tang, Hengli, author
Type of Resource: text
Genre: Journal Article
Text
Date Issued: 2017-08-11
Physical Form: computer
online resource
Extent: 1 online resource
Language(s): English
Abstract/Description: Allergy to peanuts has become a common and severe problem, especially in westernized countries. In this study, we evaluated the target and epitope specificity of the capture and detection mouse monoclonal antibodies (mAbs) used in a commercial peanut allergen detection platform. We first identified the target of these antibodies as Ara h 3 and then used an overlapping peptide array of Ara h 3 to determine the antibody-binding epitopes. Further amino acids critical for the binding via alanine substitutions at individual amino acid residues within the epitope were mapped. Finally, inhibition ELISA and inhibition immunoblotting using a recombinant Ara h 3 protein were performed to confirm these results. Surprisingly, the capture and detection mAbs showed identical binding characteristics and were presumed to represent two isolates of the same clone, a notion supported by both isoelectric focusing electrophoresis and Liquid chromatography-mass spectrometry experiments. The simultaneous binding of a pair of identical mAbs to an individual allergen such as Ara h3 is attributed to the multivalency of the analyte and has implications for developing diagnostic assays for additional multimeric allergens.
Identifier: FSU_pmch_28800361 (IID), 10.1371/journal.pone.0182935 (DOI), PMC5553815 (PMCID), 28800361 (RID), 28800361 (EID), PONE-D-17-08766 (PII)
Publication Note: This NIH-funded author manuscript originally appeared in PubMed Central at https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5553815.
Subject(s): Alanine/chemistry
Alanine/genetics
Alanine/immunology
Allergens/chemistry
Allergens/immunology
Amino Acid Sequence
Amino Acid Substitution
Animals
Antibodies, Monoclonal/biosynthesis
Antibodies, Monoclonal/chemistry
Antibodies, Monoclonal/isolation & purification
Antigens, Plant/chemistry
Antigens, Plant/genetics
Antigens, Plant/immunology
Arachis/chemistry
Arachis/immunology
Enzyme-Linked Immunosorbent Assay/standards
Epitope Mapping
Epitopes/analysis
Epitopes/chemistry
Epitopes/immunology
Gene Expression
Humans
Mice
Models, Molecular
Peanut Hypersensitivity/diagnosis
Peanut Hypersensitivity/immunology
Plant Proteins/chemistry
Plant Proteins/genetics
Plant Proteins/immunology
Protein Array Analysis
Protein Multimerization
Protein Structure, Secondary
Reagent Kits, Diagnostic
Recombinant Proteins/chemistry
Recombinant Proteins/genetics
Recombinant Proteins/immunology
Seed Storage Proteins/chemistry
Seed Storage Proteins/genetics
Seed Storage Proteins/immunology
Persistent Link to This Record: http://purl.flvc.org/fsu/fd/FSU_pmch_28800361
Owner Institution: FSU
Is Part Of: PloS one.
1932-6203
Issue: iss. 8, vol. 12

Choose the citation style.
Zhao, L., Zhao, L., Zhang, B., Robotham, J. M., Roux, K. H., & Tang, H. (2017). Identification of a common Ara h 3 epitope recognized by both the capture and the detection monoclonal antibodies in an ELISA detection kit. Plos One. Retrieved from http://purl.flvc.org/fsu/fd/FSU_pmch_28800361