Development of Indirect Competitive Enzyme-Linked Immunosorbent Assay and Lateral-Flow Immunochromatographic Strip for the Detection of Digoxin in Human Blood

Abstract

Mouse−mouse hybridoma cell lines producing stable, highly specific monoclonal antibodies with good affinity for the cardiac glycoside digoxin (DIG) were established to construct an indirect enzyme-linked immunosorbent assay and lateral-flow immunochromatographic strip to detect DIG in human blood. The hapten DIG was coupled to bovine serum albumin or chicken ovalbumin by sodium periodate oxidation. The highest sensitivity and specificity antibody had a median inhibitory concentration (IC 50 ) of 0.45 ng/mL, a linear range of detection of 0.293−0.7 ng/mL, and low cross-reactivity with several DIG analogues. The cut-off value of the lateral-flow immunochromatographic strip was 5 ng/mL when the strip was tested with human blood. The immunochromatographic lateral flow strip test provides a quick and convenient method for determining DIG in plasma which can be visually observed in only 5 min to promote rational drug use.

Sensitive Lateral Flow Immunoassay for the Residues of Imidocarb in Milk and Beef Samples

“…Three days later, after the final sprint immunization of the target mouse, its spleen was dissected out and fused with myeloma cells using PEG 2000 in a sterile ultra clean environment. The specific method was performed as previously described from our laboratory. − One week after the fusion, the supernatant from the hybridoma cell culture from 96-well microculture plates was tested using an ELISA. The wells with high antiserum titer and inhibition were screened out for cell subcloning.…”

Evaluation of digoxin-boronate ester formation through in-capillary derivatisation-large volume sample stacking-capillary zone electrophoresis

“…Digoxin determination is performed mainly using enzymelinked immunosorbent assays, based on an antigen-antibody reaction and the change in the spectroscopic signal; however, these assays have disadvantages such as the need for special kits, lack of specicity to digoxin (since the reaction of the sensor with similar structure molecules has been observed), and errors up to 10%. 9,10 Chromatographic techniques such as HPLC-MS-MS have become a popular option for digoxin analysis, with an LOQ of 0.25 ng mL −1 ; however, the cost of the equipment and analysis are a restraining factor for most laboratories, and therefore, new approaches in digoxin sensing in different matrices are needed. 11 Boronic acids (BAs) are a group of compounds that present different applications.…”

A gold-based immunochromatographic strip for the detection of sirolimus in human whole blood

“…Therefore, it would be very useful to establish an efficient and convenient method for the monitoring of sirolimus. In recent years, immunoassays have become very popular for both food 18 and drug 19 monitoring; these techniques include chemiluminescent enzyme immunoassays (CLEIAs), 20 chemiluminescence microparticle immunoassays (CMIAs), 21 colloidal gold immunochromatographic test strips 22,23 and indirect competitive enzyme-linked immunosorbent assays (ic-ELISAs). 24,25 Recently a type of chemiluminescent immunoassay using a polyclonal antibody against sirolimus has been reported.…”