An intensely luminescent polymeric lanthanide chelator for multiple fluorescence labeling of biomolecules

Tetrahedron Letters

Volumn 37, Issue 36, 1996, Pages 6483-6486

  Lamture, Jagannath B ^a   Iverson, Bonnie ^a   Hogan, Michael E ^a  

^a BAYLOR COLLEGE OF MEDICINE   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

CHELATING AGENT;

ARTICLE; DNA BINDING; DRUG SYNTHESIS; FLUORESCENCE;

EID: 0030565510     PISSN: 00404039     EISSN: None     Source Type: Journal    
DOI: 10.1016/0040-4039(96)01417-7     Document Type: Article

References (20)

1. Kricka, L. J. Nonisotopic DNA Probe Techniques, Academic Press, New York, 1992, 3.
2. a)Meares,C.F; Wensel, T.G. Acc.Chem.Res. 1984, 17, 202.
3. b)Hirschfield,T. AppLOpt, 1976, 15, 3135.
4. Hassan M.; Landon, J; Smith, D. FEBS Lett. 1979, 103, 339.
5. Lamture,J.B.; Wensel,T.G. Tetrahedron Lett. 1993, 34, 4141 and references cited therein
6. a) Wallac Inc. Publication # 1244-1126-03, Published by Offset House, Oy, Naantali, Finland, 1992.
7. b) Lovgren, T.; Hurskainen, P.; Dahlen P. Nonisotopic DNA Probe Techniques, Ed. Kricka, L. J., Academic Press, New York, 1992, 227.
8. Evangelista, R.A.; Pollak,A.; Allore,B.; Templeton, E.F. G..; Morton, R.B. ; Diamandis, E.P. Clin.Biochem. 1988, 21, 173.
9. Takalo.H.; Kankare.J.; Hanninen,E. Acta. Chem. Scancdinavica, B42, 1988, 448.
10. 5 : 478.002574 and Observed : 478.003060, at 12000 ppm resolution.
11. Miller, T. L.; Senkfor, S.I. Anal. Chem. 1992, 54, 2022.
12. A detailed investigation of DNA target labelling and fluorescence detection of hybridization to solid surfaces will be reported elsewhere.
13. 10,000 MW poty-L-lysine.HBr(15umol as lysyl units) was reacted with 2 (30μmol) in dry DMSO (1mL) and DIEA(250μmol), followed by succinylation with succinic anhydride(200μmol) and DMAP (30μmol). The resulting product was hydrolysed in 0. 1M NaOH, followed by purification via dialysis (Spectrapor-6 tubing) and Centricon-3 concentration.The degree of modification of the lysyl groups was monitored by a TNBS test for primary amines as reported by Means, G.E.; Feeney, RE. (1971), Chemical Modifications of Proteins, Holden-Day Inc. San Francisco, p217.
14. Lamture, J.B. ; Wensel, T.G. Bioconjugate Chem. 1995, 6, 88.
15. Soini, E.; Lovgren, T. CRC, Crit Rev. Anal Chem. 1987, 18, 105.
16. Extinction coefficient of the monomer has been determined gravitometrically, and that of the PLCS polymer based upon a standard yield of 20-40 3b equivalents per poly-L-lysine, as determined in 8 and assuming that the extinction coefficient of the monomer is not altered in the polymer.
17. Emission lifetime of the 3:1 PLCS:Eu(III) complex has been measured with a home-built device, based on a chopped Ar-laser (for details see Lamture, J. B.; Zhou, Z; Kumar, A.S.; Wensel, T. G. Inorganic Chem. 1995, 34, 864), generously provided by Dr. Wensel. Decay was adequately fit as the sum of two components, 0.58 and 2ms, amounting to 0.6 and 0.4 of the overall amplitude, respectively.
18. Quantum yield in solution has been measured at the absorption and emission maxima of die chelator, on a Hitachi F-2000 fluorimeter, for excitation at 337 nm. Ethidium bromide/ duplex DNA of equal absorption at 334 nm was used as a standard. Ethidium bromide, when complexed with duplex DNA, has a quantum yield of 0.8. ( source : Molecular Probes).
19. Solid surface quantum yield was obtained by measuring the luminescence of luL wet / dried microspots of the PLCS-Eu(III) and Ethidium bromide-DNA duplex deposited on polyethylene film and illuminated with a broadband long wavelength transilluminator. The luminescence densities were quantified on a BioRad 620 densitometer by comparing the yield of dried and undried microdroplets.
20. 32P-labeled oligomers at known specific activity. Quantitation was perfomed via scintillation counting. Beads per unit volume were determined on a Hausser Scientific Neubauer Hemacytometer-3520.