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. 1987 Dec;84(23):8225–8229. doi: 10.1073/pnas.84.23.8225

Monoclonal antibodies to DNA modified with cis- or trans-diamminedichloroplatinum(II).

W I Sundquist 1, S J Lippard 1, B D Stollar 1
PMCID: PMC299514  PMID: 2446320

Abstract

Murine monoclonal antibodies that bind selectively to adducts formed on DNA by the antitumor drug cis-diamminedichloroplatinum(II), cis-DDP, or to the chemotherapeutically inactive trans isomer trans-DDP were elicited by immunization with calf thymus DNA modified with either cis- or trans-DDP at ratios of bound platinum per nucleotide, (D/N)b, of 0.06-0.08. The binding of two monoclonal antibodies to cis-DDP-modified DNA was competitively inhibited (50% control) in an enzyme-linked immunosorbent assay (ELISA) by 4-6 nM concentrations (600-900 fmol) of cis-DDP bound to DNA, (D/N)b = 0.031. Similar concentrations (4-6 nM) of cis-DDP-modified poly(dG).poly(dC) also inhibited antibody binding, whereas higher concentrations (17-36 nM) of cis-DDP-modified poly[d(AG)].poly[d(TC)] were required for inhibition. Adducts formed by cis-DDP on other synthetic DNA polymers did not inhibit antibody binding to cis-DDP-DNA. The biologically active compounds [Pt(en)Cl2], [Pt(dach)Cl2], and [Pt(NH3)2(cbdca)] (carboplatin) (where en is ethylenediamine, dach is 1,2-diaminocyclohexane, and cbdca is cyclobutane-1,1-dicarboxylate) all formed antibody-detectable adducts on DNA, whereas the inactive platinum complexes trans-DDP and [Pt(dien)Cl]Cl (dien, diethylenetriamine) did not. The monoclonal antibodies therefore recognize a bifunctional Pt-DNA adduct with cis stereochemistry in which platinum is coordinated by two adjacent guanines or, to a lesser degree, by adjacent adenine and guanine. A monoclonal antibody raised against trans-DDP-DNA was competitively inhibited in an ELISA by 40 nM trans-DDP bound to DNA, (D/N)b = 0.022. This antibody crossreacted with unmodified, denatured DNA. Its binding to trans-DDP-DNA was selectively inhibited by trans-DDP-modified poly[d(GT)].poly[d(CA)] (50% inhibition at 1 nM bound trans-DDP). The recognition of cis- or trans-DDP-modified DNAs by monoclonal antibodies thus parallels the known modes of DNA binding of these compounds and may correlate with their biological activities.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Butour J. L., Johnson N. P. Chemical reactivity of monofunctional platinum-DNA adducts. Biochemistry. 1986 Aug 12;25(16):4534–4539. doi: 10.1021/bi00364a012. [DOI] [PubMed] [Google Scholar]
  2. Eastman A., Barry M. A. Interaction of trans-diamminedichloroplatinum(II) with DNA: formation of monofunctional adducts and their reaction with glutathione. Biochemistry. 1987 Jun 16;26(12):3303–3307. doi: 10.1021/bi00386a009. [DOI] [PubMed] [Google Scholar]
  3. Fichtinger-Schepman A. M., Lohman P. H., Berends F., Reedijk J., van Oosterom A. T. Interactions of the antitumour drug cisplatin with DNA in vitro and in vivo. IARC Sci Publ. 1986;(78):83–99. [PubMed] [Google Scholar]
  4. Fichtinger-Schepman A. M., van der Veer J. L., den Hartog J. H., Lohman P. H., Reedijk J. Adducts of the antitumor drug cis-diamminedichloroplatinum(II) with DNA: formation, identification, and quantitation. Biochemistry. 1985 Jan 29;24(3):707–713. doi: 10.1021/bi00324a025. [DOI] [PubMed] [Google Scholar]
  5. Galfrè G., Milstein C. Preparation of monoclonal antibodies: strategies and procedures. Methods Enzymol. 1981;73(Pt B):3–46. doi: 10.1016/0076-6879(81)73054-4. [DOI] [PubMed] [Google Scholar]
  6. Knox R. J., Friedlos F., Lydall D. A., Roberts J. J. Mechanism of cytotoxicity of anticancer platinum drugs: evidence that cis-diamminedichloroplatinum(II) and cis-diammine-(1,1-cyclobutanedicarboxylato)platinum(II) differ only in the kinetics of their interaction with DNA. Cancer Res. 1986 Apr;46(4 Pt 2):1972–1979. [PubMed] [Google Scholar]
  7. Lepre C. A., Strothkamp K. G., Lippard S. J. Synthesis and 1H NMR spectroscopic characterization of trans-[Pt(NH3)2[d(ApGpGpCpCpT)-N7-A(1),N7-G(3)]]. Biochemistry. 1987 Sep 8;26(18):5651–5657. doi: 10.1021/bi00392a011. [DOI] [PubMed] [Google Scholar]
  8. Malfoy B., Hartmann B., Macquet J. P., Leng M. Immunochemical studies of DNA modified by cis-dichlorodiammineplatinum(II) in vivo and in vitro. Cancer Res. 1981 Oct;41(10):4127–4131. [PubMed] [Google Scholar]
  9. Pinto A. L., Lippard S. J. Binding of the antitumor drug cis-diamminedichloroplatinum(II) (cisplatin) to DNA. Biochim Biophys Acta. 1985;780(3):167–180. doi: 10.1016/0304-419x(85)90001-0. [DOI] [PubMed] [Google Scholar]
  10. Pinto A. L., Lippard S. J. Sequence-dependent termination of in vitro DNA synthesis by cis- and trans-diamminedichloroplatinum (II). Proc Natl Acad Sci U S A. 1985 Jul;82(14):4616–4619. doi: 10.1073/pnas.82.14.4616. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Plooy A. C., Fichtinger-Schepman A. M., Schutte H. H., van Dijk M., Lohman P. H. The quantitative detection of various Pt-DNA-adducts in Chinese hamster ovary cells treated with cisplatin: application of immunochemical techniques. Carcinogenesis. 1985 Apr;6(4):561–566. doi: 10.1093/carcin/6.4.561. [DOI] [PubMed] [Google Scholar]
  12. Poirier M. C., Lippard S. J., Zwelling L. A., Ushay H. M., Kerrigan D., Thill C. C., Santella R. M., Grunberger D., Yuspa S. H. Antibodies elicited against cis-diamminedichloroplatinum(II)-modified DNA are specific for cis-diamminedichloroplatinum(II)-DNA adducts formed in vivo and in vitro. Proc Natl Acad Sci U S A. 1982 Nov;79(21):6443–6447. doi: 10.1073/pnas.79.21.6443. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Rahmouni A., Malinge J. M., Schwartz A., Leng M. Comparison between poly(dG-dC).poly(dG-dC) and DNA modified by cis-diamminedichloroplatinum (II): immunological and spectroscopic studies. J Biomol Struct Dyn. 1985 Oct;3(2):363–375. doi: 10.1080/07391102.1985.10508423. [DOI] [PubMed] [Google Scholar]
  14. Reed E., Yuspa S. H., Zwelling L. A., Ozols R. F., Poirier M. C. Quantitation of cis-diamminedichloroplatinum II (cisplatin)-DNA-intrastrand adducts in testicular and ovarian cancer patients receiving cisplatin chemotherapy. J Clin Invest. 1986 Feb;77(2):545–550. doi: 10.1172/JCI112335. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Stone P. J., Kelman A. D., Sinex F. M. Resolution of alpha, beta and gamma DNA of Saccharomyces cerevisiae with the antitumor drug cis-Pt (NH3)2CL2. Evidence for preferential drug binding by GpG sequences of DNA. J Mol Biol. 1976 Jul 15;104(4):793–801. doi: 10.1016/0022-2836(76)90182-0. [DOI] [PubMed] [Google Scholar]
  16. Strickland P. T., Boyle J. M. Immunoassay of carcinogen-modified DNA. Prog Nucleic Acid Res Mol Biol. 1984;31:1–58. doi: 10.1016/s0079-6603(08)60374-8. [DOI] [PubMed] [Google Scholar]
  17. Sundquist W. I., Lippard S. J., Stollar B. D. Binding of cis- and trans-diamminedichloroplatinum(II) to deoxyribonucleic acid exposes nucleosides as measured immunochemically with anti-nucleoside antibodies. Biochemistry. 1986 Apr 8;25(7):1520–1524. doi: 10.1021/bi00355a008. [DOI] [PubMed] [Google Scholar]
  18. Wells R. D., Larson J. E., Grant R. C., Shortle B. E., Cantor C. R. Physicochemical studies on polydeoxyribonucleotides containing defined repeating nucleotide sequences. J Mol Biol. 1970 Dec 28;54(3):465–497. doi: 10.1016/0022-2836(70)90121-x. [DOI] [PubMed] [Google Scholar]

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