Quantitative sandwich ELISA for the determination of tropomyosin from crustaceans in foods.
The ubiquitous muscle protein tropomyosin has been identified as the major shrimp allergen and is suggested to be a cross-reacting allergen. Previously, only a few methods for the detection of tropomyosin in food have been published. A quantitative sandwich enzyme-linked immunosorbent assay (ELISA) for the detection of tropomyosin from crustaceans in foods has been developed and validated. A polyclonal rabbit antitropomyosin capture antibody and the biotinylated conjugate of the same antibody for detection were the basis for the ELISA, which was specific for crustaceans. The ELISA was able to quantitate tropomyosin in various food matrixes, had a detection limit of 1 microg/g, and cross-reacted to some extent with cockroach. Recoveries ranged from 63 to 120%, and the intra and interassay coefficients of variation were <6 and <14%, respectively.
Werner
Marianne T
MT
National Veterinary Institute, P.O. Box 8156 Dep., N-0033 Oslo, Norway. marianne.werner@vetinst.no
Faeste
Christiane K
CK
Egaas
Eliann
E
D000485:Allergens / Q000032:analysis* / Q000737:chemistry; D000818:Animals; D000906:Antibodies; D019921:Biotinylation; D003445:Crustacea / Q000737:chemistry*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D011817:Rabbits; D012680:Sensitivity and Specificity; D016415:Sequence Alignment; D012758:Shellfish / Q000032:analysis*; D014335:Tropomyosin / Q000032:analysis* / Q000737:chemistry / Q000276:immunology
D000485:Allergens; D000906:Antibodies; D014335:Tropomyosin