A Class-Selective Immunoassay for Sulfonamides Residue Detection in Milk Using a Superior Polyclonal Antibody with Broad Specificity and Highly Uniform Affinity

Abstract

The development of multianalyte immunoassays with an emphasis on food safety has attracted increasing interest, due to its high target throughput, short detection time, reduced sample consumption, and low overall cost. In this study, a superior polyclonal antibody (pAb) against sulfonamides (SAs) was raised by using a bioconjugate of bovine serum albumin with a rationally designed hapten 4-[(4-aminophenyl) sulfonyl-amino]-2-methoxybenzoic acid (SA10-X). The results showed that the pAb could recognize 19 SAs with 50% inhibition (IC50) below 100 µg L−1 and a recognition profile for SAs containing, either a five-atom ring or a six-atom ring, with highly uniform affinity. A three-dimensional quantitative structure-activity relationship analysis indicated that the electrostatic features of SAs play a considerably important role, during recognition with pAb than stereochemical effects. Skimmed milk samples were directly diluted five times before analysis. After optimization, the limit of detection for sulfamonomethoxine, sulfamethoxazole, sulfaquinoxaline, sulfadimethoxine, and sulfamethazine were 1.00, 1.25, 2.95, 3.35, and 6.10 µg L−1, respectively. The average recoveries for these 5 SAs were 72.0–107.5% with coefficients of variation less than 14.1%. The established method, based on pAb, with broad specificity and uniform affinity, offered a simple, sensitive, and high-throughput screening tool for the detection of multi-SAs in milk samples.

Onsite/on‐field analysis of pesticide and veterinary drug residues by a state‐of‐art technology: A review

“…There are five types of labeled immunoassays: radioimmunoassay (RIA), fluorescence immunoassay (FIA), chemiluminescence immunoassay (CLIA), enzyme-linked immunoassay (ELISA), and bio-barcode immunoassay, and they are employed as biosensing analytical tools [194,195]. Among them, the enzyme-based [197] Chlorpyrifos and fenthion Tangerine juices 0.20 ± 0.04 and 0.50 ± 0.06 μg/L, respectively [189] Clindamycin and lincomycin Animal tissues 1.8 and 6.8 μg/kg, respectively [198] Penicillin G, amoxicillin, ampicillin, penicillin V, cloxacillin, oxacillin, cloxacillin, carbenicillin, dicloxacillin, and methicillin, nafcillin Milk 0.7−9.3 μg/kg [199] Permethrin, aroclor1248, and aroclor1254 Soil/sediment and house dust 1.5−14.3 μg/kg [200] Sulfonamides (sulfamonomethoxine, sulfamethoxazole, sulfaquinoxaline, sulfadimethoxine, and sulfamethazine) Milk 1.00−6.10 μg/L [201] chloramphenicol, avermectin, tetracycline, and streptomycin Food 0.106−0.134 ng/mL [202] Ethylene thiourea Soil, tomato, and potato 0.201 ng/mL [203] Trimethoprim, diaveridine, brodimoprim, and ormetoprim, baquiloprim…”

Rapid Multi-Residue Detection Methods for Pesticides and Veterinary Drugs

“…This is because of the required simple chemical structure comparisons at the two-dimensional (2D) level when the haptens are designed. With the development of computer-assisted molecular modeling, the influences of the hapten structures and spacers on steric and electronic properties can be investigated and evaluated at the three-dimensional (3D) level [21]. The potential binding sites and modes of antibody-analyte recognition can be predicted by molecular docking and computational chemistry techniques [22].…”

Preparation and Characterization of Monoclonal Antibodies with High Affinity and Broad Class Specificity against Zearalenone and Its Major Metabolites

“…In recent years scholars have researched antigen design and synthesis to broaden the recognition spectrum of antibodies. Thus far, the following three approaches have been used to obtain a broad spectrum of specific antibodies via antigen design and synthesis: (1) The development of a general structural immunogen, with the general structure of the analytes as the detection target, such as aflatoxins (AFs) [31], sulfonamides (SAs) [32], organophosphorus (OPs) [33], and fluoroquinolones (FQs) [34]. (2) The development of a multi-hapten immunogen by simultaneously coupling several different haptens to one carrier protein, such as avermectins (AVMs) [35], microcystins (MCs) [36], and pesticides, including chlorpyrifos, triazophos, carbofuran, and parathion methyl [29].…”

“…Besides, each B lymphocyte produces a specific type of antibody molecule [45]. Therefore, many immunoassays can be optimized to achieve a higher sensitivity using heterologous coating antigens [32,36]. A heterologous indirect non-competitive ELISA (inELISA) and a heterologous indirect competitive ELISA (icELISA) were employed to select the cell fusion mice and the positive hybridioma cells.…”