Quantification of the major brown shrimp allergen Pen a 1 (tropomyosin) by a monoclonal antibody-based sandwich ELISA

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 cr...

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Published in:	Journal of allergy and clinical immunology Vol. 100; no. 2; pp. 229 - 234
Main Authors:	Jeoung a, Byeoung-Ju, Reese a, Gerald, Hauck b, Peter, Oliver b, Jerry B., Daul a, Carolyn B., Lehrer a, Samuel B.
Format:	Journal Article
Language:	English
Published:	New York, NY Mosby, Inc 01-08-1997 Elsevier
Subjects:	Allergens - analysis Allergens - immunology Animals Antibodies, Monoclonal Antigens, allergens Biological and medical sciences Brachyura - chemistry Brachyura - immunology Cross Reactions cross-reactivity crustacea Decapoda (Crustacea) - chemistry Decapoda (Crustacea) - immunology Enzyme-Linked Immunosorbent Assay - methods Female Food allergy Fundamental and applied biological sciences. Psychology Fundamental immunology Immediate hypersensitivity. Allergy. Anaphylaxis, etc Immunobiology Mice Mice, Inbred BALB C Nephropidae - chemistry Nephropidae - immunology Pen a 1 monoclonal antibodies quantification Reference Standards Regression Analysis sandwich ELISA Sensitivity and Specificity shrimp Tropomyosin - analysis Tropomyosin - immunology Pen a 1 monoclonal antibodies Food allergy SDS-PAGE shrimp quantification CV TBS sandwich ELISA crustacea cross-reactivity TMC mAb Tropomyosin Brachyura Contractile protein Penaeus aztecus Quantization Prawn Monoclonal antibody Enzyme immunoassay Macrura Crustacea Crab Lobster Arthropoda Animal Cross reaction Decapoda Invertebrata ELISA assay Food Allergen
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Summary:	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.)
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0091-6749 1097-6825
DOI:	10.1016/S0091-6749(97)70229-X