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.

[1]  R. Blake,et al.  Enzyme immunoassay to determine heavy metals using antibodies to specific metal-EDTA complexes: optimization and validation of an immunoassay for soluble indium. , 1994, Analytical biochemistry.

[2]  K. Wagatsuma,et al.  Role of a Binary Metallic Modifier in the Determination of Cadmium in Graphite Furnace Atomic Absorption Spectrometry , 2010, Analytical sciences : the international journal of the Japan Society for Analytical Chemistry.

[3]  L. Thuy,et al.  Multielemental characterization of airborne particulate matter collected in Bucharest and Tokushima by inductively coupled plasma mass spectrometry and inductively coupled plasma atomic emission spectrometry. , 2010, Analytical sciences : the international journal of the Japan Society for Analytical Chemistry.

[4]  J. Turnlund.  Molybdenum metabolism and requirements in humans. , 2002, Metal ions in biological systems.

[5]  N. Kang,et al.  Determination of Cd and Cr in an ABS candidate reference material by instrumental neutron activation analysis. , 2008, Applied radiation and isotopes : including data, instrumentation and methods for use in agriculture, industry and medicine.

[6]  Yan Zhang,et al.  Determination of chloramphenicol residues in milk by enzyme-linked immunosorbent assay: improvement by biotin-streptavidin-amplified system. , 2010, Journal of agricultural and food chemistry.

[7]  J. Barbet,et al.  Preliminary study of the metal binding site of an anti-DTPA-indium antibody by equilibrium binding immunoassays and immobilized metal ion affinity chromatography. , 1995, Bioconjugate chemistry.

[8]  Dangqin Jin,et al.  Preliminary results on the determination of ultratrace amounts of cadmium in tea samples using a flow injection on-line solid phase extraction separation and preconcentration technique to couple with a sequential injection hydride generation atomic fluorescence spectrometry. , 2005, Talanta.

[9]  F. Bittner,et al.  Cell biology of molybdenum. , 2006, Biochimica et biophysica acta.

[10]  E. Stiefel.  The biogeochemistry of molybdenum and tungsten. , 2002, Metal ions in biological systems.

[11]  D. Wylie,et al.  Metal Binding Properties of a Monoclonal Antibody Directed toward Metal-Chelate Complexes* , 1996, The Journal of Biological Chemistry.

[12]  R. Mendel,et al.  Molybdoenzymes and molybdenum cofactor in plants , 2002 .

[13]  C. Meares,et al.  Conjugation of antibodies with bifunctional chelating agents: isothiocyanate and bromoacetamide reagents, methods of analysis, and subsequent addition of metal ions. , 1984, Analytical biochemistry.

[14]  R. Mendel,et al.  Biosynthesis and molecular biology of the molybdenum cofactor (Moco). , 2002, Metal ions in biological systems.