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

doi:10.1007/s11783-011-0349-8

Front Envir Sci Eng Chin

2011, Vol. 5

Issue (3) : 409-416 https://doi.org/10.1007/s11783-011-0349-8

RESEARCH ARTICLE

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

Huan HE¹, Bo TANG², Cheng SUN¹(

), Shaogui YANG¹, Weijuan ZHENG²(

), Zichun HUA²

1. The State Key Laboratory of Pollution Control and Resource Reuse, School of the Environment, Nanjing University, Nanjing 210093, China; 2. The State Key Laboratory of Pharmaceutical Biotechnology, School of Life Science, Nanjing University, Nanjing 210093, China

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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 UV-Vis. 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^-1–10³ 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 0–10³ 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.

Keywords cadmium hapten monoclonal antibody enzyme-linked immunosorbent assay (ELISA)

Corresponding Author(s): SUN Cheng,Email:envidean@nju.edu.cn; ZHENG Weijuan,Email:wjzheng@nju.edu.cn

Issue Date: 05 September 2011

Cite this article:

Cheng SUN,Shaogui YANG,Weijuan ZHENG, et al. Preparation of hapten-specific monoclonal antibody for cadmium and its ELISA application to aqueous samples[J]. Front Envir Sci Eng Chin, 2011, 5(3): 409-416.

URL:

https://academic.hep.com.cn/fese/EN/10.1007/s11783-011-0349-8
https://academic.hep.com.cn/fese/EN/Y2011/V5/I3/409

Fig.1 FT-IR spectra of IEDTA and Cd-IEDTA (molar ratio was 1∶1)

Fig.2 UV-Vis spectra of (a) BAS, Cd-IEDTA, and Cd-IEDTA-BSA; (b) KLH, Cd-IEDTA, and Cd-IEDTA-KLH

Tab.1 Reactivity of serum from BALB/c mice injected with Cd-IEDTA-KLH to IEDTA-BSA and Cd-IEDTA-BSA

Fig.3 Representative results of hybirdoma screening by competitive ELISA

Fig.4 Inhibition of different concentration of EDTA to supernatant of antibody 3E2

Fig.5 Standard inhibition curve for cadmium obtained by monoclonal antibody 3E2. The plots were fitted by the sigmoidal curve

Tab.2 Cross-reactivity determined by competitive ELISA with other metal ions

Tab.3 Precision of intra- and inter-assay of indirect competitive ELISA for cadmium (= 5)

Tab.4 Recovery of the immunoassay for cadmium in ultrapure, tap and Taihu Lake water ( = 5)

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