New haptens and antibodies for ractopamine

Wang, Zhanhui; Liu, Meixuan; Shi, Weimin; Li, Chenglong; Zhang, Suxia; Shen, Jianzhong

doi:10.1016/j.foodchem.2015.03.043

Cited by 43 publications

(29 citation statements)

References 14 publications

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“…S1a), demonstrating the conjugation between RAC and BSA. When compared with previous methods used for conjugating RAC and BSA (Wang et al, 2015;Buakeaw et al, 2016), the conjugate molar ratios of the various methods were reported as 7:1 and 8:1. Unfortunately, the drawbacks of the previous conjugation methods are that they required complicated steps, many reagents, strong organic solvents and nitrogen gas for solvent evaporation.…”

Section: Conjugation Of Rac and Protein Carriermentioning

confidence: 92%

“…2c). Furthermore, the strip test with the RAC-BSA prepared via the Mannich reaction (RAC-BSA (1) ) was compared to the strip test with the RAC-BSA prepared by the previous method (Wang et al, 2015;Buakeaw et al, 2016) (RAC-BSA (2) ) using the same antibody and concentration of the RAC-BSAs. After loading the running buffer, the color intensity appeared on the test line immobilized by RAC-BSA (1) (Fig.…”

Section: Conjugation Of Rac and Protein Carriermentioning

confidence: 99%

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Novel ractopamine–protein carrier conjugation and its application to the lateral flow strip test for ractopamine detection in animal feed

Preechakasedkit

Ngamrojanavanich

Khongchareonporn

et al. 2019

J. Zhejiang Univ. Sci. B

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In this work, a novel conjugate of ractopamine and bovine serum albumin (RAC-BSA) has been developed via the Mannich reaction, with a mole coupling ratio for RAC-BSA of 9:1. The proposed conjugation method provides a simple and one-step method with the use of fewer reagents compared with other conjugation methods for competitive immunoassays. RAC-BSA conjugation was used to fabricate a competitive lateral flow strip test for RAC detection in animal feed. For sample preparation, RAC was spiked in swine feed purchased from the local markets in Thailand, and methanol and running buffer at a volume ratio of 10:90 was used as extraction buffer. The procedures for sample preparation were completed within 25 min. Under optimal conditions, the limit of detection (LOD), assessed by the naked eye within 5 min, was found to be 1 ng/g. A semi-quantitative analysis was also conducted using a smart phone and computer software, with a linearity of 0.075-0.750 ng/g, calculated LOD of 0.10 ng/g, calculated limit of quantitation of 0.33 ng/g, and good correlation of 0.992. The recoveries were found in the range of 96.4%-103.7% with a relative standard deviation of 2.5%-3.6% for intra-and inter-assays. Comparison of the results obtained by the strip test with those obtained by enzyme-linked immunosorbent assay had a good agreement in terms of accuracy. Furthermore, this strip test exhibited highly specific RAC detection without cross reactivity with related compounds. Therefore, the RAC-BSA conjugation via the Mannich reaction can be accepted as a one-step and easy conjugation method and applied to the competitive lateral flow strip test.

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Section: Conjugation Of Rac and Protein Carriermentioning

confidence: 92%

Section: Conjugation Of Rac and Protein Carriermentioning

confidence: 99%

Novel ractopamine–protein carrier conjugation and its application to the lateral flow strip test for ractopamine detection in animal feed

Preechakasedkit

Ngamrojanavanich

Khongchareonporn

et al. 2019

J. Zhejiang Univ. Sci. B

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“…They usually have to be modified in order to be coupled with carrier proteins called haptens. Therefore, appropriate design and synthesis of haptens is the key step in antibody production, which could influence the affinity and specificity of the antibody as well as its performance in an immunoassay [18]. In immunology it is widely accepted that antibodies preferentially recognize the part of molecule that is the farthest away from the hapten.…”

Section: Traditional Antibodiesmentioning

confidence: 99%

Immunochemical detection of emerging organic contaminants in environmental waters

Zhang

Zeng

Liu

2017

TrAC Trends in Analytical Chemistry

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“…And one of the most well-known strategy was employing heterologous coating antigen to improve detection sensitivity [7,8]. However, how the sensitivity was affected by coating antigen was poorly understood.…”

Section: Introductionmentioning

confidence: 99%

Spectra analysis of coating antigen: A possible explanation for difference in anti-AFB1 polyclonal antibody sensitivity

Yang

Liu

Wang

et al. 2016

Journal of Molecular Structure

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For the detection of small hapten molecules, indirect competitive enzyme-linked immunosorbent assay (icELISA) is a preferred method. However, diverse coating antigen might bring different antiserum titer and sensitivity for the identical antiserum. In the present study, four AFB1-protein (aflatoxin B1-carrier protein) conjugates were prepared by activated ester method (AFB1O-BSA/AFB1O-OVA) and mannich method (AFB1-cBSA/AFB1-cOVA), and then applied as coating antigen for titer and sensitivity detection of the identical antiserum obtained from rabbit immunized by AFB1-KLH. Afterwards, the ultraviolet-visible, fluorescence and far-ultraviolet circular dichroism (far-UV CD) spectra were recorded for understanding the difference in titer and sensitivity obtained. Results revealed that AFB1O-BSA/AFB1O-OVA showed a strong intrinsic fluorescence band centered at 450 nm that originated from the emission of AFB1, which differed from AFB1-cBSA/AFB1-cOVA, while the decrease of α-helical and increase of β-sheet in AFB1-cBSA was the most remarkable. This indicated that the better sensitivity obtained by using AFB1O-BSA as coating antigen might be caused by its extended structure, because such structure affect the binding between AFB1 and antibody. The study might offer structural information for understanding the titer and sensitivity difference caused by coating antigen.

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New haptens and antibodies for ractopamine

Cited by 43 publications

References 14 publications

Novel ractopamine–protein carrier conjugation and its application to the lateral flow strip test for ractopamine detection in animal feed

Novel ractopamine–protein carrier conjugation and its application to the lateral flow strip test for ractopamine detection in animal feed

Immunochemical detection of emerging organic contaminants in environmental waters

Spectra analysis of coating antigen: A possible explanation for difference in anti-AFB1 polyclonal antibody sensitivity

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