![]() To date, few studies have been conducted on the blocking effect using different microplate types. To prevent NSB, blocking is an essential step to saturate the unoccupied sites on the solid phase. For example, NSB of antibodies in sera has been reported by several ELISA studies. First, during each assay step, any substances may adsorb to the solid phase due to non-specific binding (NSB), causing a high background reading or false immunosignal. There are two major factors that affect the accuracy and reproducibility of ELISA. Reproducibility can be regarded as precision, which is a measurement of the variation in samples in the same assay (within the same run) or different assays (from day to day or from different experimenters). Accuracy is the degree of closeness of the determined value to the nominal or known true value under prescribed conditions. ( A) Rabbit anti-P Hb pAb and mouse anti-P Hb mAb was applied as the capture and detection antibody, respectively ( B) mouse anti-P Hb mAb and rabbit anti-P Hb pAb was applied as the capture and detection antibody, respectively ( C) rabbit anti-P Hb pAb and biotinylated mouse anti-P Hb mAb was applied as the capture and detection antibody, respectively.Īccuracy and reproducibility are two of the criteria during assay validation. In indirect sELISA, detection antibodies can also be labeled with biotin, which can further interact with enzyme-labeled avidins, such as streptavidin-horseradish peroxidase (HRP) conjugate ( Figure 1C). It should be noted that the use of secondary antibodies may lead to cross-reaction, which is defined as any unexpected interaction between a particular antibody and those non-specific antigens. In the indirect format ( Figure 1), the unlabeled detection antibody can be identified by the labeled secondary antibody ( Figure 1A,B). However, this labeling process could be time-consuming and expensive. In the direct format, the enzymes-, fluorophores-, or nanoparticles- conjugated detection antibody enables immunosignal recognition. Monoclonal (mAb) or polyclonal antibody (pAb) can be used for the capture or detection antibody in sELISA, which can be performed either directly or indirectly. In general, sandwich ELISA (sELISA) is one of the formats that can be commercialized due to its standardized quality control and simple operation. comprises one of the world’s largest markets. The global ELISA market was valued at about USD 1.6 billion in 2018 and is projected to increase significantly at a compound annual growth rate of 5.5% from 2019 to 2028. In addition, ELISA has been widely used in hospitals, clinical laboratories, pharmaceutical companies, and research organizations. In 2019, ELISA accounted for 61% of the total global food safety testing market, and it is a dominant technique for the detection of food adulterants. Among different methods for the surveillance of food fraud, enzyme-linked immunosorbent assay (ELISA) is widely applied due to its advantages of sensitivity, rapidity, selectivity, reproducibility, economy, efficiency, and easiness to handle without complex instruments. Recently, many studies have reported the potential increase of food fraud due to the COVID-19 pandemic. Globally, it is estimated that food fraud affects approximately 10% of food products and leads to a loss of approximately USD 10–15 billion each year. Food fraud includes a wide range of deliberate fraudulent acts to foods such as substitution, addition, tampering, dilution, counterfeiting, or misrepresentation of foods or food ingredients, which may cause potential health risks. ![]()
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