Monoclonal-based enzyme-linked immunosorbent assay and immunochromatographic assay for enrofloxacin in biological matrices

Abstract

Enrofloxacin has been increasingly used in veterinary medicine to treat microbial infections. A simple and reliable analytical method for this drug is required. The current determination by high performance liquid chromatography (HPLC) is sensitive but labor-intensive. This paper reports an enzyme-linked immunosorbent assay (ELISA) using a monoclonal antibody (MAb) and the development of a rapid test kit based on immunochromatography. The detection limits using the ELISA were 10 ppb for chicken liver and muscle, and 1 ppb for cattle milk, respectively. The mean recovery values were 77.3-96.0% for chicken liver, 72.4-92.0% for chicken muscle and 84.0-99.0% for cattle milk. The detection limits using the kit were ca. 100 ppb for chicken muscle and ca. 10 ppb for cattle milk, respectively. All ELISA results for assay of chicken liver, chicken muscle and cattle milk were confirmed using HPLC which is used as the routine assay. The HPLC (x) and ELISA (y) results showed close correlation for chicken liver (y = 8.7 + 0.85x, r2 = 0.99, n = 25), chicken muscle (y = -3.9 + 0.94x, r2 = 0.98, n = 25) and cattle milk (y = 18.4 + 0.92x, r2 = 0.99, n = 25).

Preparation of high-affinity rabbit monoclonal antibodies for ciprofloxacin and development of an indirect competitive ELISA for residues in milk

“…Numerous studies have been reported on developing ELISA methods for the detection of FQs, including CPFX (Mellgren and Sternesjö, 1998), enrofloxacin (Mellgren and Sternesjö, 1998;Watanabe et al, 2002), norfloxacin (Watanabe et al, 2002;Huet et al, 2006), pefloxacin (Mellgren and Sternesjö, 1998;Huet et al, 2006), and sarafloxacin (Lu et al, 2006). Moreover, Bucknall et al (2003) reported results concerning a polyclonal antibody against CPFX with a limit of detection (LOD) of 4 ng/ml (Bucknall et al, 2003).…”

Detection of Enrofloxacin and Its Metabolite Ciprofloxacin Using Gold Nanoparticles and Laser Desorption/Ionization Time-of-Flight Mass Spectrometry

“…The sensitivity of our method is comparable to ELISA and electrophoresis, and may not have an advantage over routine analytical methods such as HPLC, LC/MS, and ELISA in terms of sensitivity. [12][13][14][15][16][17][18][19][20][21][22][23] However, our approach is advantageous over these methods since it has a short assay time, a simple experimental protocol, and low background signals. We believe that our strategy has wide potential for use as a detection method for small molecules that have corresponding antibodies or capturing agents.…”

“…[6][7][8] A number of countries have therefore regulated the use of these two FQs for animal bred for food production; for example, the European Union and the United States have set maximum residue limits (MRLs) in edible tissues, milk, and eggs, and the use of these two drugs is restricted for laying hens. [9][10][11] Along with tightening up the regulations on FQ use, various analytical methods have been developed for determining or screening FQ residues, including high-performance liquid chromatography (HPLC), [12][13][14] liquid chromatography-mass spectrometry (LC/MS), 15,16 enzyme-linked immunosorbent assays (ELISA), [17][18][19][20] and capillary electrophoresis. [21][22][23] Although these methods are routinely used with high sensitivity, general use of these methods is sometimes hampered by the need for sophisticated and time-consuming processes.…”

Sensitive immunoassay for simultaneous determination of tetrabromobisphenol A bis(2-hydroxyethyl) ether and tetrabromobisphenol A mono(hydroxyethyl) ether: An effective and reliable strategy to estimate the typical tetrabromobisphenol A derivative and byproduct in aquatic environments

“…In fact, the farthest parts of the M3 molecular structure coupling site (hapten and the carrier protein) are the same as TBBPA DHEE and TBBPA MHEE. According to the rule in immunology (Bucknall et al, 2003;Watanabe et al, 2002;Zhang et al, 2013), antibodies elicited from haptenic conjugates showed a preferential recognition to the molecule part that was farthest from coupling site. In this study, TBBPA DHEE and TBBPA MHEE (the structures of both chemicals are shown in Fig.…”