Development and application of one-step ELISA for the detection of neomycin in milk

Abstract

A sensitive enzyme-linked immunosorbent assay (ELISA) for the detection of neomycin (NEO) in milk was developed. Two conjugates were synthesized as the immunogen and coating antigen. Four BALB/c female mice were immunised and the antisera were screened by indirect competitive ELISA (icELISA). The icELISA was further optimised using different coating methods, pH values, ionic strengths of assay buffer and reaction times. After optimised, the indirect competitive ELISA (icELISA) could be finished in 85min with an IC 50 value of 0.7490.05 ng/mL. One-step indirect competitive ELISA (icELISA) exhibited similar sensitivity with an IC 50 value of 0.7390.03 ng/mL, which was sensitive enough for NEO detection and could be finished in 55 min. No cross-reactivity of the antibody was observed with other aminoglycosides based on icELISA, indicating that the antibody is highly specific for neomycin. Milk samples were further analyzed with recovery ranging from 85 to 110% at spiked level of 0.25Á10 ng/mL, and the detection limit was 0.08 ng/mL.

Development of an ELISA for nitrazepam based on a monoclonal antibody

“…Stop solution (50 µL/well) was then added and the plate was measured using an ELISA plate reader instrument at 450 nm (Xu et al, 2011).…”

Development of generic immunoassay for the detection of a series of aminoglycosides with 6′-OH group for the treatment of genetic diseases in biological samples

“…Over the last few decades many analytical and bioanalytical assays have been suggested for the qualitative and quantitative analysis of aminoglycosides that are in clinical, veterinary and agricultural use to treat bacterial infections. These methods were applied for the detection of drug residues in a wide range of biological derived matrices such as cell tissues [15], serum [16], milk [17], eggs [15,18], and honey [18]. Chemical methodologies included the application of gas chromatography (GC) [19], thin layer chromatography (TLC) [20], high-performance liquid chromatography (HPLC) [21] and capillary electrophoresis (CE) [22].…”

Development of a monoclonal antibody assay and a lateral flow strip test for the detection of paromomycin residues in food matrices

“…The immunogens and coating antigens were prepared with the glutaraldehyde (GDA) and carbodiimide (EDC) coupling methods reported in previous studies (Isanga et al, 2016;Xu et al, 2011;Xu et al, 2014), as briefly described below.…”

“…However, paromomycin, like other aminoglycosides, cannot be measured by UV absorption because it lacks a chromophore and has poor chromatographic properties on reversed-phase liquid chromatography (Faten et al, 2015). Chromatographic methods are also expensive and require time-consuming and laborious sample pretreatment steps, sophisticated laboratory equipment, and technical personnel (Li et al, 2016;Lourenco, Barbosa, & Pinto, 2011;Xu et al, 2011). Therefore, alternative methods for determining paromomycin residues in foods must be developed.…”