Rapid determination of phenylethanolamine A in biological samples by enzyme-linked immunosorbent assay and lateral-flow immunoassay

Abstract

Phenylethanolamine A (PA) is a β-adrenergic agonist, which was first used in animal husbandry as a growth promoter in China in 2010. In this study, a monoclonal-antibody (mAb)-based indirect competitive enzyme-linked immunosorbent assay (icELISA) and lateral-flow immunoassay (LFA) for the detection of PA in swine urine and pork were developed. The immunogen was prepared by linking PA hapten with carrier protein via a diazotization method. The IC50 value of the optimized icELISA was 0.44 ng mL(-1). The limits of detection of the icELISA for PA in swine urine and pork were 0.13 ng mL(-1) and 0.39 ng g(-1), respectively. The recoveries of PA from spiked swine urine and pork were in the range 82.0-107.4 % and 81.8-113.3%, respectively, with the coefficients of variation in the range 4.1-16.2% and 1.2-6.3%, respectively. The mAbs had negligible cross reactivity with 10 other β-agonists. In contrast, the LFA had a cut-off level of 5 ng mL(-1) (g) in swine urine and pork, and the results could be achieved within 5 min. Ten blind samples of swine urine were analyzed simultaneously by icELISA, LFA, and ultra-high-performance liquid chromatography-tandem mass spectrometry, and the results of the three methods agreed well. Therefore, the combination of two immunoassays provides an effective and rapid screening method for detection of PA residues in biological samples.

Cationic conjugated polymer-based FRET aptasensor for label-free and ultrasensitive ractopamine detection

“… 8 However, their cost of equipment, highly demanding testing environment, and high time consumption limit their extensive application under limited experimental conditions. 9 Therefore, a series of simple and rapid analytical methods for RAC detection, such as enzyme-linked immunosorbent assay (ELISA), 10 chemiluminescence 11 and electrochemistry, 12 have been introduced for RAC analysis. It is worth noting that these technologies commonly require a labelling process, which not only increases the cost of analysis but might also reduce the specificity and affinity of the RAC-related probes.…”

Development of a highly sensitive and specific monoclonal antibody based enzyme‐linked immunosorbent assay for the detection of a new β‐agonist, phenylethanolamine A, in food samples

“…From Table , it can be seen that except one ultra‐sensitive pAb‐ELISA developed in our laboratory, the sensitivity of our mAb‐ELISA is 1.3 to 39 times higher (e.g. IC 50 value is 1.3 to 39 times lower) than those in other ELISAs, demonstrating the high sensitivity of our mAb‐ELISA. In addition, our mAb‐based ELISA also displays other advantages such as high specificity and unlimited supply of antibody by cell culture for hybridoma.…”

Current Advances in Immunoassays for the Detection of β2-Agonists

“…Indirect competitive ELISAs have been developed to detect β 2 -agonists, including phenylethanolamine A [40], ractopamine [41], zilpaterol [42], clenbuterol [43,44] and salbutamol [45,46] in various types of samples. In these assays, the β 2 -agonist coated on the microtiter plate competes with β 2 -agonist in samples for binding to the primary anti-β 2 -agonist antibody.…”

“…It has an intense color and no development process is needed for visualization. Colloidal gold has been used by multiple laboratories to label an antibody against β 2 -agonist and develop LFIA for detecting target β 2 -agonist in biological samples, such as clenbuterol in urine and pork muscle [50,51], salbutamol in urine, meat and milk [52,53], phenylethanolamine A in urine and pork [40,54] and ractopamine in animal urine, meat, liver and feed [50,[55][56][57][58]. Huang et al [59] used bacteria as a carrier of gold nanoparticles to construct bacteria-Au-antibody probe in LFIA to detect clenbuterol.…”