Preparation of hapten-specific monoclonal antibody for cadmium and its ELISA application to aqueous samples

Abstract

High-affinity and specific monoclonal antibodies against cadmium-ethylene diamine tetraacetic acid (EDTA) complex have been produced using the hybridoma technique. A hapten was synthesized and characterized by Fourier Transform Infrared Spectroscopy (FT-IR) and UV-Vis. Competitive enzyme-linked immunosorbent assay (ELISA) for quantitative detection of cadmium in aqueous sample was developed. The monoclonal antibody with high level of binding affinity for Cd-IEDTA-BSA and high specificity for soluble Cd-EDTA complex showed less than 0.99% cross-reactivity with other 11 metals. The limit of detection was 0.10 μg·L^-1, and the effective linear range was 10^-1–10³ μg·L^-1. The intra- and inter-assay coefficient variations were 1.5%–6.3% and 3.2%–7.4%, respectively. The spike recovery in different water samples were between 98.5% and 110.3%. The detection limit of this assay was well below the allowable concentration of cadmium (3 μg·L^-1), and the working range was wider than that of other methods which showed the range of 2.19–86.38 and 0–10³ μg·L^-1. The competitive ELISA established in this paper was sensitive and accurate in the screening of cadmium in aqueous samples. The results will lay a solid foundation for construction of an immunoassay kit for cadmium.

Keywords

cadmium / hapten / monoclonal antibody / enzyme-linked immunosorbent assay (ELISA)

References

[1] Satarug S, Moore M R. Adverse health effects of chronic exposure to low-level cadmium in foodstuffs and cigarette smoke. Environmental Health Perspectives, 2004, 112(10): 1099–1103

[2] Christen K, Renner R. How green is ethanol? | Pretreatment lowers desalination costs | News Briefs: Sick-stream diagnosis | Cadmium linked to cancer | OMB proposes to harmonize risk assessments. Environmental Science & Technology, 2006, 40(6): 1722–1724

[3] Meranger J C, Subramanian K S, Chalifoux C. A national survey for cadmium, chromium, copper, lead, zinc, calcium, and magnesium in Canadian drinking water supplies. Environmental Science & Technology, 1979, 13(6): 707–711

[4] Komarnicki G J K. Lead and cadmium in indoor air and the urban environment. Environmental Pollution, 2005, 136(1): 47–61

[5] John M K, Chuah H H, VanLaerhoven C J. Cadmium contamination of soil and its uptake by oats. Environmental Science & Technology, 1972, 6(6): 555–557

[6] Amweg E L, Weston D P. Whole-sediment toxicity identification evaluation tools for pyrethroid insecticides: I. Piperonyl butoxide addition. Environmental Toxicology and Chemistry /SETAC, 2007, 26(11): 2389–2396

[7] Cabrera C, Lorenzo M L, Gallego C, Lopez M C, Lillo E. Cadmium levels in food and feed crops, determined by electrothermal atomic absorption spectrometry. Journal of Agricultural and Food Chemistry, 1992, 40(9): 1631–1633

[8] Cabrera C, Lorenzo M L, Gallego C, Lopez M C, Lillo E. Cadmium contamination levels in seafood determined by electrothermal atomic absorption spectrometry after microwave dissolution. Journal of Agricultural and Food Chemistry, 1994, 42(1): 126–128

[9] Díaz S, Martín-González A, Carlos Gutiérrez J. Evaluation of heavy metal acute toxicity and bioaccumulation in soil ciliated protozoa. Environment International, 2006, 32(6): 711–717

[10] Wang S T, Gui W J, Guo Y R, Zhu G N. Preparation of a multi-hapten antigen and broad specificity polyclonal antibodies for a multiple pesticide immunoassay. Analytica Chimica Acta, 2007, 587(2): 287–292

[11] Wilson R. Microtitre plate enzyme amplified immunoassay for thyroid stimulating hormone. Analytica Chimica Acta, 1992, 269(2): 301–306

[12] Chuang J C, Emon J M V, Durnford J, Thomas K. Development and evaluation of an enzyme-linked immunosorbent assay (ELISA) method for the measurement of 2,4-dichlorophenoxyacetic acid in human urine. Talanta, 2005, 67(3): 658–666

[13] Reardan D T, Meares C F, Goodwin D A, McTigue M, David G S, Stone M R, Leung J P, Bartholomew R M, Frincke J M. Antibodies against metal chelates. Nature, 1985, 316(6025): 265–268

[14] Chakrabarti P, Hatcher F M, Blake II R C, Ladd P A, Blake D A. Enzyme immunoassay to determine heavy metals using antibodies to specific metal-EDTA complexes: optimization and validation of an immunoassay for soluble indium. Analytical Biochemistry, 1994, 217(1): 70–75

[15] Wylie D E, Lu D, Carlson L D, Carlson R, Babacan K F, Schuster S M, Wagner F W. Monoclonal antibodies specific for mercuric ions. In: Proceedings of the National Academy of Sciences of the United States of America, 1992, 89(9): 4104–4108

[16] Blake D A, Chakrabarti P, Khosraviani M, Hatcher F M, Westhoff C M, Goebel P, Wylie D E, Blake II R C 2nd. Metal binding properties of a monoclonal antibody directed toward metal-chelate complexes. The Journal of Biological Chemistry, 1996, 271(44): 27677–27685

[17] Johnson D K. A fluorescence polarization immunoassay for cadmium(II). Analytica Chimica Acta, 1999, 399(1-2): 161–172

[18] Zhu X X, Hu B S, Lou Y, Xu L N, Yang F L, Yu H N, Blake D A, Liu F Q. Characterization of monoclonal antibodies for lead-chelate complexes: applications in antibody-based assays. Journal of Agricultural and Food Chemistry,2007,55 (13), 4993–4998

[19] Johnson D K, Combs S M, Parsen J D, Jolley M E. Lead analysis by anti-chelate fluorescence polarization immunoassay. Environmental Science & Technology, 2002, 36(5): 1042–1047

[20] Levy R, Shohat L, Solomon B. Specificity of an anti-aluminium monoclonal antibody toward free and protein-bound aluminium. Journal of Inorganic Biochemistry, 1998, 69(3): 159–163

[21] Blake D A, Pavlov A R, Yu H N, Kohsraviani M, Ensley H E, Blake R C. Antibodies and antibody-based assays for hexavalent uranium. Analytica Chimica Acta, 2001, 444(1): 3–11

[22] Sarada N C, Thamaraiselvi K, Vijayalakshmi M A. Anti Zn antibodies: cross reactivity and competitive binding with heavy metals. Journal of Chromatography B, Analytical Technologies in the Biomedical and Life Sciences, 2008, 861(2): 236–239

[23] Darwish I A, Blake D A. One-step competitive immunoassay for cadmium ions: development and validation for environmental water samples. Analytical Chemistry, 2001, 73(8): 1889–1895

[24] Tawarada K, Sasaki K, Ohmuka N, Matsumoto N, Saiki H. Preparation of Anti-cadmium-EDTA complex monoclonal antibody and its binding specificity. Japan Society for Analytical Chemistry, 2003, 52(8): 583–587

[25] Zhu X X, Xu L, Lou Y, Yu H N, Li X, Blake D A, Liu F Q. Preparation of specific monoclonal antibodies (MAbs) against heavy metals: MAbs that recognize chelated cadmium ions. Journal of Agricultural and Food Chemistry, 2007, 55(19): 7648–7653

[26] Sasaki K, Oguma S, Glass T, Namiki Y, Sugiyama H, Ohmura N, Blake D A. Simple method to reduce interference from excess magnesium in cadmium immunoassays. Journal of Agricultural and Food Chemistry, 2008, 56(17): 7613–7616

[27] Kriegel A M, Soliman A S, Zhang Q, El-Ghawalby N, Ezzat F, Soultan A, Abdel-Wahab M, Fathy O, Ebidi G, Bassiouni N, Hamilton S R, Abbruzzese J L, Lacey M R, Blake D A. Serum cadmium levels in pancreatic cancer patients from the East Nile Delta region of Egypt. Environmental Health Perspectives, 2006, 114(1): 113–119

[28] He H, Liu Z Y, Yang S G, Sun C. Preparation of anti-cadmium-EDTA complex polyclonal antibody and its application for determination of cadmium in aqueous solution. Analytical Letters, 2009, 42(2): 409–424

[29] Leaist D G, Hao L. Tracer diffusion of some metal ions and Metal–EDTA complexes in aqueous sodium chloride solutions. Journal of the Chemical Society, Faraday Transactions, 1994, 90(1): 133–136

[30] Pecosk R L. Polarography of Copper in Ethylenediaminetetraacetic Acid Solutions. Analytical Chemistry, 1953, 25(4): 561–564

[31] Furukawa K, Takahashi Y, Sato H. Effect of the formation of EDTA complexes on the diffusion of metal ions in water. Geochimica et Cosmochimica Acta, 2007, 71(18): 4416–4424

[32] Saito S, Danzaka N, Hoshi S. Direct fluorescence detection of Pb²⁺ and Cd²⁺ by high-performance liquid chromatography using 1-(4-aminobenzyl)ethylenediamine-N,N,N′N′-tetraacetate as a pre-column derivatizing agent. Journal of Chromatography. A, 2006, 1104(1–2): 140–144

[33] Rinehart K, Namikoshi M, Choi B. Structure and biosynthesis of toxins from blue-green algae (cyanobacteria). Journal of Applied Phycology, 1994, 6(2): 159–176

[34] Saito S, Danzaka N, Hoshi S. Direct fluorescence detection of Pb²⁺ and Cd²⁺ by high-performance liquid chromatography using 1-(4-aminobenzyl)ethylenediamine-N,N,N′N′-tetraacetate as a pre-column derivatizing agent. Journal of Chromatography. A, 2006, 1104(1–2): 140–144

[35] Khosraviani M, Pavlov A R, Flowers G C, Blake D A. Detection of heavy metals by immunoassay: Optimization and validation of a rapid, portable assay for ionic cadmium. Environmental Science & Technology, 1998, 32(1): 137–142

[36] Blake R C II, Pavlov A R, Khosraviani M, Ensley H E, Kiefer G E, Yu H N, Li X, Blake D A. Novel monoclonal antibodies with specificity for chelated uranium(VI): isolation and binding properties. Bioconjugate Chemistry, 2004, 15(5): 1125–1136

RIGHTS & PERMISSIONS

Higher Education Press and Springer-Verlag Berlin Heidelberg