The effect of fish matrix on the enzyme‐linked immunosorbent assay of antibiotics

Abstract

Significant interference in the immunoassay of norfloxacin was observed in the presence of fish matrix. Some proteins and ions were demonstrated to contribute to the matrix effect investigated. Although the detailed mechanism is still unclear, the non-specific interaction between fish proteins and immunoglobulin G (IgG) or horseradish peroxidase (HRP) labelled IgG was assumed to be an important source of the matrix effect in immunoassays.

The matrix interference to the immunoassay of food samples: The effect of some proteins in aquatic products

“…However, it is not sure that, such a nonspecific binding has universal significance as a source of interference for other immunoassays, or it is just due to the high conformational similarity between the proteins and the antigen (parvalbumin). In our previous study (Wang, Lin, Sui, & Cao, 2013), some proteins from flatfishes were also demonstrated to interfere strongly with the enzyme-linked immunosorbent assay (ELISA) of an antibiotic (norfloxacin); moreover, they could result in significant positive bands in western blotting in the presence of antibody reagents, indicating that the fish protein-IgG interaction might play an important role in the matrix interference observed.…”

Effect of pH shifts on IgE-binding capacity and conformational structure of tropomyosin from short-neck clam (Ruditapes philippinarum)

“…The tropomyosin treated with different pH was analyzed with SDS-PAGE according to Wang's methods (Wang, Lin, Sui, & Cao, 2013). Samples (5 lg) were reduced by heating with loading buffer and loaded into each well of the SDS-PAGE gel (12% for running gel, 5% for stacking gel).…”

5-sulfosalicylic Acid Dihydrate-Based Pretreatment for the Modification of Enzyme-Linked Immunoassay of Fluoroquinolones in Fishery Products

“…SDS-PAGE and Western blotting were preformed according to the method of Wang et al [16] The polyvinylidene difluoride (PVDF) membrane on which proteins has electro-blotted was blocked in PBS (containing 1 g L −1 Tween 20 and 50 g L −1 defatted milk) overnight at 4.Then the membrane was incubated at room temperature for 2 hr with HRP-labelled goat anti-mouse IgG (1:1000v/v) diluted in PBS. The reactive bands were visualized using chemiluminescent substrates (Thermo Scientific, Rockford, IL, USA), and then recorded by Tanon180 Gel imaging system…”

“…Chemical compounds present in samples or sample extracts, such as protein, solvents, and others, might affect the binding of antibody and analytes, and it also might inhibit enzyme activity. [17] In our previous studies, [16] some fish proteins weighted about 36 kDa and 42 kDa in sample extracts might contribute much to matrix interference by non-specific interaction with immunoglobulin G (IgG) or horseradish peroxidase (HRP) labeled IgG.…”

“…According to Wang's reports, [16] a lot of proteins, from molecular weight below 14.4 kDa to about 116 kDa, could be extracted from the fishery samples by PBS. Among those proteins, the two components with molecular weight approximate 36 kDa and 42 kDa, could bind with the horseradish peroxidase (HRP) labeled IgG effectively.…”