Monoclonal-based enzyme-linked immunosorbent assay and immunochromatographic rapid assay for monensin†

Abstract

Monensin, a member of the ionophoric polyether antibiotics, is used primarily as a coccidiostat. A protein conjugate of monensin was prepared and utilized to produce monoclonal antibodies in the BALB/c-P3X63Ag8U.1 fusion system. Only one hybridoma that produces monoclonal antibody against monensin was isolated from one in 329 wells. The monoclonal antibody was used to develop quantitative assays for monensin by means of an enzyme-linked immunosorbent assay (ELISA). The detection limit was 1 ng ml-1 and the relative standard deviations were 2.1-6.3% intra-assay and 5.9-12.9% inter-assay. All ELISA results for assay of chicken plasma and cattle milk were confirmed using a bioassay to be used as the official method. The ELISA and bioassay results showed close correlations for plasma (r2 = 0.98, n = 25) and milk (r2 = 0.95, n = 25). Using the anti-monensin monoclonal antibodies produced, a rapid test kit based on the immunochromatographic method was developed. Detection limits of monensin for cattle milk, cattle plasma and chicken plasma were about 40, 40 and 160 ppb. respectively.

Pretreatment-free lateral flow enzyme immunoassay for progesterone detection in whole cows’ milk

“…The next step of the work was the selection of optimal membrane components and reagents concentration for sensitive competitive LFEIA. Five types of commercial analytical membranes with different pore size (15,10,8,5 μm) and capability to bind proteins (CNPF -lower, CNPC -higher and CNPH -the highest protein binding) were considered. A range of lateral flow assay characteristics (assay time, sensitivity etc.)…”

“…The authors used paper test strip totally covered by specific antibodies and the assay was based on the measurement of the height of colored bars formed on strips as a result of immunoreaction with the following enzyme substrate saturation. Later enzyme immunoassay was realized in conventional LFIA format with specific reagents immobilized in a form of narrow bands [8]. In enzyme LFIA the most often used label is horse radish peroxidase (HRP) [3][4][5][6], however alkaline phosphatase [8,9] and cholinesterase [10] were also employed.…”

“…Later enzyme immunoassay was realized in conventional LFIA format with specific reagents immobilized in a form of narrow bands [8]. In enzyme LFIA the most often used label is horse radish peroxidase (HRP) [3][4][5][6], however alkaline phosphatase [8,9] and cholinesterase [10] were also employed. As far as HRP is concerned, colorimetric [3][4][5][6], chemiluminescent [11,12] and electrochemical [13] detection modes were reported.…”

A one-step immunochromatographic assay for detecting ginsenosides Rb1 and Rg1

“…[12,13,14,15,16]. A variety of immunochromatographic assays have been reported for detection of sulfadimidine residues [17], monensin residue [18], and coeliac disease [19]. An immunochromatographic assay is based on a competitive immunoassay that utilizes antigen-antibody binding properties and provides rapid and sensitive detection of analyte.…”

Development of immunoassays for the detection of kanamycin in veterinary fields

“…Watanabe et al [19] reported on a monoclonal-based ELISA and an immunochromatographic assay for monitoring monensin residues in chicken plasma and cattle milk. In addition, several recent studies have reported on a colloidal gold-based immunochromatographic assay.…”