Monoclonal antibody based immunoassay: An alternative way for aquatic environmental selenium detection

Environmental concerns about human health encouraged increasing methodological interest in selenium (Se), which is an essential non-metal trace element and varies within a narrow concentration range between essential and toxic. In this study, two types of long-armed Se haptens (Se-hapten-lc-NHS) were synthesized for the first time using active ester formalization. In producing monoclonal antibodies (mAbs), the derivatization of haptenized Se at para- (meta-) and ortho-sites showed different properties. Finally, a mAb derived from hybridoma 5A5² was confirmed to be capable of establishing an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA). There was a successful quantitative determination of Se⁴⁺ with a detection range of 17 to 207 pmol mL^-1 and a limit of detection of approximately 3.9 pmol mL^-1. The mAb was found to be remarkably sensitive and specific, with no evidence of cross-reactivity with other ions. The assay was validated for four kinds of Se forms in water samples and showed satisfactory recoveries between 80 % and 108 %, with coefficients of variation of 2.1 %-11 %. The method proposed in our study offers a useful protocol for the rapid screening of Se and provides an alternative solution for the analysis of Se in aquatic environments.