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In this paper, an indirect competitive enzyme‐linked immunosorbent assay (IC‐ELISA) has been developed and optimized to detect Hg(II) in tap water and lake water based on a monoclonal antibody (mAb‐A24). Some stabilizing additives (Gelatin, bovine serum albumin [BSA], polyvinyl alcohol [PVA], and polyvinyl pyrrolidone [PVP]) and surfactant (Tween‐20) have been investigated thoroughly in the optimization process. Under the optimal condition, the 50% half maximal inhibitory concentration (IC50) and limit of detection (LOD) were 1.68 and 0.079 ng/ml, respectively. These anti‐Hg mAbs also have some affinity with methyl mercury (CH3Hg) and with no cross‐reactivity with other thirteen metal ions. The developed method has shown satisfactory recovery of Hg(II), ranged between 91% and 116%, from tap water and lake water. Therefore, this immunoassay can be used to detect trace Hg(II) in environment water.

Paper-based lateral flow immunoassays (LFIAs) are an interesting on site technique for detecting heavy metals. Pb(II) is one of the toxic heavy metals in polluted environmental water. This paper reports the synthesis and characterization of three Fe3O4-based nanoadsorbents that show different performance to Pb(II), Cu(II), Cd(II), Hg(II), As(V), Zn(II), Fe(III), Ca(II), Mg(II), Mn(II), and Al(III) when a GO or an amino group is introduced. Compared with Fe3O4, Fe3O4-GO showed higher adsorption percentage to many heavy metal ions under slightly acid condition. The negative surface on Fe3O4@NH2 had favourable interactions for four main divalent metal ion adsorption, which followed the order: Pb(II) > Cu(II) > Hg(II) > Cd(II). Combined with antibody-functionalized AuNPs showed high affinity and specificity toward Pb(II), an Fe3O4@NH2-based LFIA with five times greater sensitivity for detecting Pb(II) in lake water was developed. The cut-off value of this Fe3O4@NH2-combined LFIA method for detection Pb(II) in lake water was 20 ng/mL and the detection result for 10 ng/mL Pb(II) was obviously observable by the naked eye or a strip reader machine. This detection strategy could be readily adapted for other small molecules or biomolecules as an ultrasensitive screening tool.