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

Abstract

The elimination of matrix interference is now a task for further development and application of immunoassays. Here the effect of proteins in aquatic samples on immunoassay was investigated by western blotting and competitive indirect enzyme-labeled immunosorbent assay (ci-ELISA). Some proteins in flatfishes interacted with different antibodies, resulting in significant loss of accuracy of the immunoassays. Moreover, these matrix proteins prefer to interact with enzyme-labeled antibodies than label-free ones. Such a matrix protein-immunoglobulin interaction was validated with different fish and shrimp samples and similar results were observed, which demonstrated a universal significance of the protein-induced matrix interference to immunoassays. Besides, the strategies to prevent such interference were preliminarily discussed.

The effect of fish matrix on colloidal gold immunochromatographic assay of antibiotics: the source of interference and removal method

“…After 5-sulfosalicylic acid (2% m/v) treatment, more than 99% proteins were removed (Table 1), and correspondingly the matrix effect was observed somewhat slighter than that in the original extracts ( Figure 2). Similar to the previous study for ELISA (Wang et al, 2016), in which some matrix protein of fishery extracts were found to react with IgG (especially enzyme-labelled antibodies), here the protein-induced matrix effect may also be attributed to its non-specific binding with the gold-labelled antibodies. This protein-IgG interaction demonstrated no significant affluence on the gold-labelled antibody for its reaction with the secondary antibody on the control line, however, significantly reduced its efficiency to bind with the antigens on the test line.…”

“…Proteins contribute much of the water-soluble components in the fish crude extracts (Table 1). In previous studies, the interference of matrix proteins to ELISA has been fully confirmed by its possible competitive interaction with antibodies (Wang et al, 2016). In this study, we have verified the influence of proteins on GICA.…”

“…The protein content was measured by the Coomassie brilliant blue method (Konstantinos, Christos, & Yves-Jacques, 2015) and the protein composition was analyzed by polyacrylamide gel electrophoresis (Wang, Li, Zheng, Liu, & Lin, 2011). The lipid content was measured according to the chloroform/methanol extraction (Carla, Davide, & Carola, 2009), and removed by diatomite filtration (Wang et al, 2016). The total sugar content was measured by the vitriolic acidphenol method with reference to the specific operation described by Tatsuya et al (2005) and the sugar composition of the concentrated crude extracts was analyzed by polyacrylamide gel electrophoresis (Theodore et al, 2005).…”

“…All these tasks are considered to be closely related with the interference from the complex food components (Tiwari et al, 2010), which is usually called "matrix effect" and has been widely indicated in GICA and enzyme-linked immune sorbent assay (ELISA) of fishery products (Christiane & Christin, 2008;Tiwari et al, 2010;Wang, Lin, Sui, & Cao, 2013). Recently understanding of the matrix effect on ELISA has made significant progress, for example, some proteins have been verified as the main interference-inducing components in fishery products, and their chemical, physical and biological characteristics, as well as the action mechanism, have been clarified (Wang et al, 2016). Based on these results, new and more effective approaches have been developed for the elimination of the matrix interference by the exploitation of 5-sulfosalicylic acid dehydrate during pre-treatments (Cui, Lin, Wang, Cao, & Sui, 2015;Qu et al, 2016).…”

Isolation, characterization, and identification of proteins interfering with enzyme-linked immunosorbent assay of antibiotics in fish matrix

“…For example, serum proteins could bind to analytes or the microplate surface (12) and allergens from codfish could bind with the monoclonal anti-parvalbumin antibody (13). In our previous study (14,17), a significant interference (p<0.05) with the immunoassay of norfloxacin was observed in the presence of flatfish matrix, and two protein components with approximate molecular weight of 42 and 36 kD, respectively, were demonstrated to effectively interact with the IgG reagents used in ELISA; therefore, they were assumed to contribute to the matrix effect investigated. However, the physical, chemical, and biological properties of the proteins are still very limited.…”