Preparation of hapten-specific monoclonal antibody for cadmium and its ELISA application to aqueous samples

Abstract

High-affinity and specific monoclonal antibodies against cadmium-ethylene diamine tetraacetic acid (EDTA) complex have been produced using the hybridoma technique. A hapten was synthesized and characterized by Fourier Transform Infrared Spectroscopy (FT-IR) and UVVis. Competitive enzyme-linked immunosorbent assay (ELISA) for quantitative detection of cadmium in aqueous sample was developed. The monoclonal antibody with high level of binding affinity for Cd-IEDTA-BSA and high specificity for soluble Cd-EDTA complex showed less than 0.99% cross-reactivity with other 11 metals. The limit of detection was 0.10 μg$L -1 , and the effective linear range was 10 -1 -10 3 μg$L -1 . The intra-and inter-assay coefficient variations were 1.5%-6.3% and 3.2%-7.4%, respectively. The spike recovery in different water samples were between 98.5% and 110.3%. The detection limit of this assay was well below the allowable concentration of cadmium (3 μg$L -1 ), and the working range was wider than that of other methods which showed the range of 2.19-86.38 and 0-10 3 μg$L -1 . The competitive ELISA established in this paper was sensitive and accurate in the screening of cadmium in aqueous samples. The results will lay a solid foundation for construction of an immunoassay kit for cadmium.

Use of brightness wavelet transformation for automated analysis of serum metallothioneins‐ and zinc‐containing proteins by Western blots to subclassify childhood solid tumours

“…Chicken anti‐zinc antibodies were prepared by HENA, Prague, Czech Republic. Two hens were immunised by Zn‐keyhole limpet hemocyanin complex according to He et al . From the egg yolk the IgY fraction with reactivity to Zn‐keyhole limpet hemocyanin was obtained.…”

Antibody developments for metal ions and their applications

“…The most used variant of ELISA for metal ions quantification is the indirect competitive ELISA (ic-ELISA), which a second enzyme-labelled antibody (anti-species antibody) introduced to produce a chromogenic or fluorogenic signal. Almost all heavy metal ions that the specific antibodies developed were ever quantified by ic-ELISA, and these ic-ELISAs enzyme-substrate systems include HRP-TMB (He et al, 2011;Liu et al, 2009;Zhu et al, 2019), HRP-OPD (Gao et al, 2012) and ALP-pNPP (Mandappa et al, 2012).…”

Development of a chemiluminescent enzyme immunoassay to detect cadmium in cereal samples

“…In addition, the pretreatment of the samples is time-consuming, and the digestion process often requires large amounts of strong acids that can harm the environment. By contrast, immunoassays, primarily relying on the specific reaction between the antibody and the antigen, exhibit some unique characteristics such as being rapid, simple to perform, cost-effective and friendly to the environment, which enable them to satisfy the demands of high-throughput and on-site screens (He et al, 2011;Zhou et al, 2011). Arai, Nakamura, Yoshida, and Tawarada (2016), Song, Zou, Zhu, Liu, and Kuang (2018) and Xing et al (2014) developed immunochromatographic strips for semiquantitative detection of cadmium.…”