Development of a polyclonal antibody based immunoassay for detection of molybdenum levels in environmental water

An indirect competitive ELISA (icELISA) based on anti-mice polyclonal antibody has been developed for the determination of molybdenum (Mo) levels in environmental water. For this reason, isothiocyanate method was employed to synthesize the artificial antigen of Mo-ITCBE-BSA, and Balb/C mice were used to prepare the polyclonal antibody. Based on the square matrix titration, an icELISA method was developed. The dynamic range was from 0.16 to 78 ng/mL, with LOD and IC₅₀ value of 0.08 ng/mL and 3.2 ng/mL, respectively. Except for cross-reactivity to Co²⁺ (36.3%) and Hg²⁺ (42.5%), negligible cross-reactivity to other metal ions was observed. Under optimization dilution folds, the recoveries of Mo²⁺ were in the range of 95.3-117.5%, 88.7-113.5% and 97.5-104.2% for tap water, river water and lake water, respectively. Therefore, this assay has the potential to be incorporated into a quantitative monitoring program for the rapid screening of molybdenum levels in water.