[Show abstract][Hide abstract] ABSTRACT: We examined the preparation method of the calibration standard for an ELISA kit for measuring crustacean protein in food products. The initial extract of the calibration standard was stabilized by addition of protease inhibitor to the extracting solution and heating at 100 degrees C for 10 min. In accordance with the preparation procedure for calibrators, we prepared 3 lots of initial extract of calibration standard and analyzed the protein concentration and SDS-electrophoretic pattern. Single bands at 160, 41, 37 kDa and 4 bands in the range of 16-20 kDa were observed on SDS-PAGE. The range of protein concentration of the initial extract of the calibration standard was 2.74-4.10 mg/mL.
No preview · Article · Jun 2010 · Journal of the Food Hygienic Society of Japan (Shokuhin Eiseigaku Zasshi)
[Show abstract][Hide abstract] ABSTRACT: The labeling of 5 major allergenic ingredients (egg, milk, wheat, buckwheat, and peanut) is mandatory in Japan, and 2 series of enzyme-linked immunosorbent assay (ELISA) kits have been established as official screening methods. However, these official methods have not provided the necessary sensitivity, due in part to poor extraction efficiency. To address this need, 2 novel ELISA kits have been developed: the FASTKIT ELISA Ver. II Series and the FASPEK Allergenic Substances Detection Kit. The new kit systems use an improved extraction buffer that can extract insoluble proteins produced by processing and feature new antibodies that bind to the denatured proteins extracted with the new extraction buffer. The analytical performances of the 2 new ELISA kit series were evaluated in an interlaboratory study. Ten laboratories participated in the study and determined the major allergenic ingredients contained in 5 types of model processed food. The 2 ELISAs displayed fairly good reproducibility and sufficient recovery.
No preview · Article · Oct 2006 · Journal of AOAC International