Quantification of the major brown shrimp allergen Pen a 1 (tropomyosin) by a monoclonal antibody-based sandwich ELISA
| Autor: | B.-J. Jeoung, Samuel B. Lehrer, Gerald Reese, C.B. Daul, P Hauck, J B Oliver |
|---|---|
| Rok vydání: | 1997 |
| Předmět: |
animal structures
genetic structures Brachyura Immunology Enzyme-Linked Immunosorbent Assay Tropomyosin Cross Reactions medicine.disease_cause Sensitivity and Specificity Cross-reactivity Epitope Mice Allergen Decapoda medicine Animals Immunology and Allergy Penaeus Mice Inbred BALB C Chromatography biology fungi Antibodies Monoclonal Allergens Reference Standards biology.organism_classification Molecular biology Nephropidae Shrimp Biotinylation Prawn Regression Analysis Female |
| Zdroj: | Journal of Allergy and Clinical Immunology. 100:229-234 |
| ISSN: | 0091-6749 |
| DOI: | 10.1016/s0091-6749(97)70229-x |
| Popis: | Background: Among 13 allergens found in extracts of cooked brown shrimp ( Penaeus aztecus) the 36 kd muscle protein tropomyosin has been identified as the only major shrimp allergen (Pen a 1). Cross-reacting molecules with similar molecular weights were detected in other crustacea species such as crab, lobster, and crawfish. Because Pen a 1 and Pen a 1-like allergens are important in crustacea allergy, the aim of this study was to develop a monoclonal antibody (mAb)–based sandwich ELISA to quantify Pen a 1 and to evaluate Pen a 1 levels in four commercial shrimp, crab, and lobster extracts. Methods: Two Pen a 1-specific mAbs with different epitope specificities were selected. ELISA plates coated with captured mAb 3.2 were incubated with samples containing Pen a 1. Bound Pen a 1 was detected by a combination of biotinylated mAb 4.9.5 and alkaline phosphatase–labeled streptavidin. Results: The optimized sandwich ELISA could detect Pen a 1 concentrations ranging from 4 to 125 ng/ml. Four commercial shrimp extracts demonstrated a 40-fold difference in Pen a 1 levels (24 to 920 μg/ml). Crab and lobster extracts contained detectable levels of Pen a 1-like proteins. No reactivity to cockroach, house dust mite, oyster, codfish, or peanut extracts was detected, which indicates that the developed assay is crustacea-specific. Conclusion: A sensitive sandwich assay was developed to quantify Pen a 1. This assay will be helpful to standardize shrimp extracts in regard to the content of the major allergen, Pen a 1, and to study cross-reactivities among and evaluate occupational exposure to different crustacea species. (J Allergy Clin Immunol 1997;100:229-34.) |
| Databáze: | OpenAIRE |
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