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

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          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 (IC 50) 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.

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          Fast screening methods to detect antibiotic residues in food samples

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            Immunochemical techniques for multianalyte analysis of chemical residues in food and the environment: A review

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              Development of a monoclonal antibody-based broad-specificity ELISA for fluoroquinolone antibiotics in foods and molecular modeling studies of cross-reactive compounds.

              A competitive indirect enzyme-linked immunosorbent assay (ciELISA) using monoclonal antibodies (Mabs) having broad specificity for fluoroquinolone (FQ) antibiotics is described. Four FQs, ciprofloxacin (CIP), enrofloxacin (ENR), norfloxacin (NOR), and ofloxacin (OFL), were conjugated to bovine serum albumin for immunogens and to ovalbumin for coating antigens. A Mab C4A9H1 raised against the CIP hapten exhibited high cross-reactivity (35-100%) with 12 of 14 FQs and detected these FQs in a ciELISA below their maximum residue levels (MRLs) with good sensitivity at 50% binding inhibition (IC50). The quantitative structure-activity relationship (QSAR) between Mab C4A9H1 and various FQs by comparative molecular field analysis (CoMFA) showed a high predictive ability with a cross-validation q2 value of 0.866. Using a simple purification process and the broad-specificity ciELISA adapted for analysis of FQs in chicken muscle, chicken liver, honey, shrimp, and whole egg samples demonstrated recoveries of 60-93% for CIP, ENR, NOR, OFL, flumequine, and danofloxacin.
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                Author and article information

                Contributors
                Role: Academic Editor
                Role: Academic Editor
                Journal
                Molecules
                Molecules
                molecules
                Molecules
                MDPI
                1420-3049
                26 January 2019
                February 2019
                : 24
                : 3
                : 443
                Affiliations
                [1 ]Beijing Advanced Innovation Center for Food Nutrition and Human Health, College of Veterinary Medicine, China Agricultural University, Beijing Key Laboratory of Detection Technology for Animal-Derived Food Safety, Beijing Laboratory of Food Quality and Safety, Beijing 100193, China; mylichenglong@ 123456cau.edu.cn (C.L.); lxscau@ 123456163.com (X.L.); huijuanyang@ 123456cau.edu.cn (H.Y.); wenkai@ 123456cau.edu.cn (K.W.); laura12390@ 123456163.com (Y.C.); chaolicau2018@ 123456gmail.com (C.L.); weiyuwang2018@ 123456gmail.com (W.W.); shiwm@ 123456cau.edu.cn (W.S.); suxia@ 123456cau.edu.cn (S.Z.); wangzhanhui@ 123456cau.edu.cn (Z.W.)
                [2 ]Henan Animal Health Supervision Institute, Zhengzhou 450008, China; liyonghan1020@ 123456126.com
                [3 ]College of Veterinary Medicine, Qingdao Agricultural University, Qingdao 266109, China; liangxiao4000@ 123456163.com
                Author notes
                [* ]Correspondence: yu51422396@ 123456126.com ; Tel.: +86-10-62734565; Fax: +86-10-62731032
                Author information
                https://orcid.org/0000-0003-3043-7230
                https://orcid.org/0000-0002-0167-9559
                Article
                molecules-24-00443
                10.3390/molecules24030443
                6384663
                30691168
                59609be9-a54a-4e7b-b0a5-e56e82c03071
                © 2019 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                : 13 December 2018
                : 24 January 2019
                Categories
                Article

                milk safety,multi-sulfonamides,novel hapten,polyclonal antibody,broad specificity,highly uniform affinity

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