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 quan-titative 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.

Automated summary

Environmental concerns have led to increased interest in selenium (Se) as its concentration range has narrow boundaries between essential and toxic levels. This study synthesized two types of long-armed Se haptens using active ester formalization and demonstrated their different properties in derivatization for producing monoclonal antibodies (mAbs). An mAb derived from hybridoma 5A52 proved to be sensitive and specific, successfully establishing an indirect competitive enzyme-linked immunosorbent assay for the quantitative determination of Se4+ in water samples.