Development and validation of an indirect competitive enzyme linked-immunosorbent assay for the determination of potentially allergenic sesame (Sesamum indicum) in food.
This study was designed to develop an indirect competitive enzyme linked-immunosorbent assay (ELISA) to detect traces of sesame in food. Antibodies against sesame were prepared by immunizing a hen with a protein extract of white, peeled sesame. The ELISA did not show any cross-reactivity with 12 of 13 food ingredients tested, only for chocolate was a low cross-reactivity of 0.7% observed. To eliminate matrix effects, sesame protein standard solutions were prepared by diluting the sesame extract with blank food matrix (1:20 diluted with PBS). Recovery of sesame protein in food samples (crisp toasts, snacks, and rolls) spiked with different sesame protein concentrations ranged from 85% to 120%, with the exception of multigrain crisp toast, resulting in too high recoveries (117%-160%) and whole grain bread, yielding too low recoveries (70%-85%). In crisp bread, cracker, cereals, and snacks the limit of detection (LOD) was found to be 5 microg of sesame protein/g of food, in fresh breads and rolls, the LOD was 11 microg of sesame protein/g of food.
Husain
Fatima Tazeen
FT
Department of Analytical and Food Chemistry, University of Vienna, Wahringer Strasse 38, A-1090 Vienna, Austria.
Bretbacher
Ines Elisabeth
IE
Nemes
Albert
A
Cichna-Markl
Margit
M
D052179:Antigens, Plant / Q000032:analysis* / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005504:Food Analysis; D057230:Limit of Detection; D010940:Plant Proteins / Q000032:analysis* / Q000276:immunology; D031685:Sesamum / Q000737:chemistry / Q000276:immunology*
D052179:Antigens, Plant; D010940:Plant Proteins