Download PDF

Science China Life Sciences

, Volume 54, Issue 9, pp 813–821 | Cite as

Synthesis and characteristics of a novel artificial hapten using the copper mercaptide of penicillenic acid from penicillin G for immunoassay of heavy metal ions

Open Access
Research Papers
First Online:
Received:
Accepted:

Abstract

In this paper, we describe the synthesis of a novel copper ion hapten using the copper mercaptide of penicillenic acid (CMPA) derived from penicillin. Results from tests with immune rabbits indicate that: (i) A new antigen synthesized with CMPA has good stability and is safe for immunizing animals with no toxic phenomena being found in animal experiments; (ii) the immunogenic antigen (CMPA-BSA) can stimulate the immune system to produce specific antibodies with high titrations, up to 150000; and (iii) antibodies in antisera showed higher affinity to OVA-GSH-CuCl than OVA-GSH, which indicates that the antibodies have specific affinity towards copper ions. These results confirm that the novel hapten and relevant antigen for copper ion have been successfully synthesized, giving progress towards an immunoassay for copper ions in environmental and food samples.

Keywords

immunoassay copper ion hapten environment food sample 
Download to read the full article text

References

  1. 1.
    Blake D A, Jones R M, Darwish I A, et al. Antibody-based sensors for heavy metal ions. Biosens Bioelectron, 2001, 16: 799–809, 11679258, 10.1016/S0956-5663(01)00223-8, 1:CAS:528:DC%2BD3MXnslems7s%3DPubMedCrossRefGoogle Scholar
  2. 2.
    Devi M, Lu D, Carlson L D, et al. Inhibition of acetylcholinesterase activity in the central nervous system of the red swamp crayfish, Procambarus clarkia, by mercury, cadmium, and lead. Bull Environ Contamin Toxicol, 1995, 55: 746–750, 10.1007/BF00203762, 1:CAS:528:DyaK2MXotFShur0%3DCrossRefGoogle Scholar
  3. 3.
    Ireland M P. Heavy metal sources—uptake and distribution. In: Dillon H K, Ho M H, eds. Biological Monitoring of Heavy Metals. New York: Wiley, 1991. 263Google Scholar
  4. 4.
    Avitsin A P, Zhavoronkov A A, Rish M A, et al. Human Microelementoses. Moscow: Meditsina, 1991Google Scholar
  5. 5.
    Gismers M J, Hueso D, Procopio J R, et al. Ion-selective carbon paste electrode based on tetraethyl thiuram disulfide for copper and mercury. Anal Chim Acta, 2004, 524: 347–353, 10.1016/j.aca.2004.03.098CrossRefGoogle Scholar
  6. 6.
    Blake D A, Blake II R C, Khosraviani M, et al. Immunoassays for metal ions. Anal Chim Acta, 1998, 376: 13–19, 10.1016/S0003-2670(98)00437-1, 1:CAS:528:DyaK1cXntVCqtbk%3DCrossRefGoogle Scholar
  7. 7.
    Ye Q Y, Li Y, Jiang Y, et al. Determination of trace cadmium in rice by flow injection on-line filterless precipitation dissolution preconcentration coupled with flame atomic absorption spectrometry. J Agricult Food Chem, 2003, 51: 2111–2114, 10.1021/jf025945+, 1:CAS:528:DC%2BD3sXitVOltrs%3DCrossRefGoogle Scholar
  8. 8.
    Allen L B, Sittonen P H, Thompson H C, et al. Methods for the determination of arsenic, cadmium, copper, lead, and tin in sucrose, corn syrups, and high-fructose corn syrups by inductively coupled plasma atomic emission spectrometry. J Agricult Food Chem, 1997, 45: 162–165, 10.1021/jf960376p, 1:CAS:528:DyaK2sXitVKqsg%3D%3DCrossRefGoogle Scholar
  9. 9.
    Shtoyko T, Maghasi A T, Richardson J N, et al. Spectroelectrochemical sensing based on attenuated total internal reflectance stripping voltammetry, 1: Determination of lead and cadmium. Anal Chem, 2003, 75: 4585–4590, 14632068, 10.1021/ac034281f, 1:CAS:528:DC%2BD3sXlvVShu74%3DPubMedCrossRefGoogle Scholar
  10. 10.
    Ronald K C, Roth J, Berson, Y. The agony and the ecstasy. J Clin Invest, 2004, 114: 1051–1054CrossRefGoogle Scholar
  11. 11.
    Marco M P, Gee S, Hammock B D, et al. Immunochemical techniques for environmental analysis I. Immunosensors. Tren Anal Chem, 1995, 14: 341–350, 1:CAS:528:DyaK2MXnt1ansrY%3DGoogle Scholar
  12. 12.
    Blake D A, Dawson G N, Chakravarti P, et al. Environmental immunochemical methods: Perspectives and applications. Washington DC: American Chemistry Society, 1996Google Scholar
  13. 13.
    Blake D A, Chakrabarti P, Hatcher F M, et al. Metal binding properties of a monoclonal antibody directed toward metal-chelate complex. J Biol Chem, 1996, 271: 27677–27685, 8910359, 10.1074/jbc.271.44.27677, 1:CAS:528:DyaK28XmvVWltb8%3DPubMedCrossRefGoogle Scholar
  14. 14.
    Lerner R A, Benkovic S J, Schultz P G. At the crossroads of chemistry and immunology catalytic antibodies. Science, 1991, 252: 659–667, 2024118, 10.1126/science.2024118, 1:CAS:528:DyaK3MXltVyqtb0%3DPubMedCrossRefGoogle Scholar
  15. 15.
    Love R A, Villafranca J E, Aust R M, et al. How the anti-(metal chelate) antibody CHA255 is specific for the metal ion of its antigen: X-ray structures for two Fab’/hapten complexes with different metals in the chelate. Biochemistry, 1993, 32: 10950–10959, 8218161, 10.1021/bi00092a004, 1:CAS:528:DyaK3sXmtVWns7o%3DPubMedCrossRefGoogle Scholar
  16. 16.
    Wylie D E, Carlson L D, Carlson R, et al. Detection of mercuric ions in water by ELISA with a mercury specific antibody. Anal Biochem, 1991, 194: 381–387, 1862940, 10.1016/0003-2697(91)90245-O, 1:CAS:528:DyaK3MXitFCjsbo%3DPubMedCrossRefGoogle Scholar
  17. 17.
    Wylie D E, Lu D, Carlson L D, et al. Immunology, monoclonal antibodies specific for mercuric ions. Proc Natl Acad Sci USA, 1992, 89: 4104–4108, 1570337, 10.1073/pnas.89.9.4104, 1:CAS:528:DyaK38XktVGktrk%3DPubMedPubMedCentralCrossRefGoogle Scholar
  18. 18.
    Meares C F, Mccall M J, Reardan D T, et al. Conjugation of antibodies with bifunctional chelating agents: isothiocyanate and bromoacetamide reagents, methods of analysis, and subsequent addition of metal ions. Anal Biochem, 1984, 142: 68–78, 6440451, 10.1016/0003-2697(84)90517-7, 1:CAS:528:DyaL2MXltVyrsg%3D%3DPubMedCrossRefGoogle Scholar
  19. 19.
    Meares C F. Chelating agents for the binding of metal ions to antibodies. Int J Rad Appl Instru B, 1986, 13: 311, 1:CAS:528:DyaL2sXjtV2lug%3D%3DCrossRefGoogle Scholar
  20. 20.
    Moi M K, Meares C F, Mccall M J, et al. Copper chelates as probes of biological systems: stable copper complexes with a macrocyclic bifunctional chelating agent. Anal Biochem, 1985, 148: 249–253, 4037304, 10.1016/0003-2697(85)90653-0, 1:CAS:528:DyaL2MXltlyjsLs%3DPubMedCrossRefGoogle Scholar
  21. 21.
    Reardan D T, Meares C F, Goodwin D A, et al. Antibodies against metal chelates. Nature, 1985, 316: 265–268, 3927170, 10.1038/316265a0, 1:CAS:528:DyaL2MXltVSqtbc%3DPubMedCrossRefGoogle Scholar
  22. 22.
    Mukkala V M, Mikola H, Hemmil I, et al. The synthesis and use of activated N-benzyl derivatives of diethylenetriaminetetraacetic acids: Alternative reagents for labeling of antibodies with metal ions. Anal Biochem, 1989, 176: 319–325, 2500868, 10.1016/0003-2697(89)90316-3, 1:CAS:528:DyaL1MXktVelsbY%3DPubMedCrossRefGoogle Scholar
  23. 23.
    Blake D A, Jones R M, Blakeii R C, et al. Antibody-based sensors for heavy metal ions. Biosens Bioelectron, 2001, 16: 799–809, 11679258, 10.1016/S0956-5663(01)00223-8, 1:CAS:528:DC%2BD3MXnslems7s%3DPubMedCrossRefGoogle Scholar
  24. 24.
    Blake R C, Delehanty J B, Khosraviani M, et al. Allosteric binding properties of a monoclonal antibody and its Fab fragment. Biochem istry, 2003, 42: 497–508, 10.1021/bi0267339, 1:CAS:528:DC%2BD38XpsF2gt7k%3DCrossRefGoogle Scholar
  25. 25.
    Chakrabarti P, Hatcher F M, Blake R C, et al. Enzyme immunoassay to determine heavy metals using antibodies to specific metal EDTA complexes: optimization and validation of an immunoassay for soluble indium. Anal Biochem, 1994, 217: 70–75, 8203740, 10.1006/abio.1994.1084, 1:CAS:528:DyaK2cXhsFKjsbw%3DPubMedCrossRefGoogle Scholar
  26. 26.
    Delehanty J B, Jones R M, Bishop T C, et al. Identification of important residues in metal-chelate recognition by monoclonal anti-bodies. Biochemistry, 2003, 42: 14173–14183, 14640685, 10.1021/bi034839d, 1:CAS:528:DC%2BD3sXovVOitb8%3DPubMedCrossRefGoogle Scholar
  27. 27.
    Khosraviani M, Pavlov A R, Flowers G C, et al. Detection of heavy metals by immunoassay: optimization and validation of a rapid, portable assay for ionic cadmium. Environ Sci Technol, 1998, 32: 137–142, 10.1021/es9703943, 1:CAS:528:DyaK2sXnsFOlsrs%3DCrossRefGoogle Scholar
  28. 28.
    Khosraviani M, Blake R C, Pavlov A R, et al. Binding properties of a monoclonal antibody directed toward lead chelate complexes. Bioconjugate Chem, 2000, 11: 267–277, 10.1021/bc9901548, 1:CAS:528:DC%2BD3cXhslymsro%3DCrossRefGoogle Scholar
  29. 29.
    Blake D A, Pavlov A R, Yu H N, et al. Antibodies and antibody-based assays for hexavalent uranium. Anal Chim Acta, 2001, 444: 3–11, 10.1016/S0003-2670(01)01151-5, 1:CAS:528:DC%2BD3MXnt1SjsrY%3DCrossRefGoogle Scholar
  30. 30.
    Maro M P, Gee S, Hammock B D. Immunochemical techniques for environmental analysis II. Antibody production and immunoassay development. Tren Anal Chem, 1995, 14: 415–418Google Scholar
  31. 31.
    Xie P, Tao X, Xu W, et al. Mercuric mercaptide of penicillenic acid, a novel hapten for relevant immunoassay, synthesized from penicillin. J Immun Meth, 2010, 353: 1–7, 10.1016/j.jim.2009.12.005, 1:CAS:528:DC%2BC3cXitValtbg%3DCrossRefGoogle Scholar
  32. 32.
    Smith J W G, Grey G E, Patel V J, et al. The spectrophotometric determination of amipicillin. Analyst, 1967, 92: 247–252, 6046816, 10.1039/an9679200247, 1:CAS:528:DyaF2sXktVGks70%3DPubMedCrossRefGoogle Scholar
  33. 33.
    Bundgaard Ilver K. A new spectrophotometric method for the determination of Penicillins. J Pharm, 1972, 24: 790–794, 10.1111/j.2042-7158.1972.tb08884.xCrossRefGoogle Scholar
  34. 34.
    He H, Liu Z Y, Yang S G, et al. Preparation of anti-cadium-EDTA complex polyclonal antibody and its application for determination of cadmium in aqueous solution. Anal Lett, 2009, 42: 409–424, 10.1080/00032710802514832, 1:CAS:528:DC%2BD1MXjvVOis7w%3DCrossRefGoogle Scholar
  35. 35.
    de Weck A L, Eisen H N. Some immunochemical properties of penicillenic acid. J Exp Med, 1960, 112: 1227–1247, 10.1084/jem.112.6.1227PubMedCentralCrossRefGoogle Scholar
  36. 36.
    Carpenter F H, Turner R A, Vigeneaud V. Benzylpenicillenic acid as an intermediate in the synthesis of benzylpenicillin (penicillin acid). J Biol Chem, 1948, 893–905Google Scholar
  37. 37.
    Singh K V, Kaur J, Varshney G C, et al. Synthesis and characterization of hapten-protein conjugates for antibody production against small molecules. Bioconjugate Chem, 2004, 15: 168–173, 10.1021/bc034158v, 1:CAS:528:DC%2BD3sXpvFKhurg%3DCrossRefGoogle Scholar

Copyright information

© The Author(s) 2011

This article is published under license to BioMed Central Ltd. Open Access This article is distributed under the terms of the Creative Commons Attribution License which permits any use, distribution and reproduction in any medium, provided the original author(s) and source are credited.

Authors and Affiliations

  1. 1.Laboratory of Analytical Biochemistry & Bio-separation, Key Laboratory of Microbiology of Ministry of Education of China, School of Life Science and BiotechnologyShanghai Jiao Tong UniversityShanghaiChina
  2. 2.School of Agriculture and BiologyShanghai Jiao Tong UniversityShanghaiChina

Personalised recommendations