Development of a monoclonal antibody-based sandwich ELISA for the detection of ovalbumin in foods. (1)
The major allergens in egg white include ovalbumin (OVA), ovomucoid, ovotransferrin and lysozyme, with OVA accounting for 54% of egg white protein. Most foods are processed in various conditions, such as baking and boiling. These processing conditions can change the solubility, structure and bioactivity of allergen proteins. Various ELISA-based methods have been developed for the detection of several food allergens.
MAb17 as the capture antibody and mAb15-HRP as the detection antibody were chosen. In addition, the two generated mAbs, mAb15 and mAb17, were identified to be IgG1 and k light chain. The limit of detection (LOD) of OVA was determined to be 0.51 ng/mL, and the linear dynamic range was from 1.95 to 500 ng/mL.
Reproducibility showed that the variation coefficients ranged from 4.82 to 8.63% for intra-assay, and from 8.54 to 13.9% for inter-assay. Recovery studies showed that the recoveries ranged from 85.6 to 111.2% for the intra-assay. In the inter-assay, the recoveries varied from 87.3 to 115.2%.
The result was positive when the S/N >2.1. Seven foods showed positive results. Four types of food samples gave negative results. It can be seen that duck egg and goose egg showed cross-reactivity. However, pigeon egg and quail egg did not show cross-reactivity.
