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Proceedings Paper

Time-resolved fluorescence study of interaction of the monoclonal anticoproporphyrin antibodies and (Pt-)coproporphyrin
Author(s): Oleg I. Lobanov; Alexander P. Savitsky; Mary Nelen; Marina V. Demcheva; Anatoly K. Yatsimirsky; Jouko E. Korppi-Tommola
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Paper Abstract

Mechanisms of ligand binding by monoclonal anti-coproporphyrin antibodies are studied by steady-state and time-resolved fluorescence spectroscopy by use of a picosecond laser system. The antibodies quench the coproporphyrin (CP) fluorescence, but the CP fluorescence spectra show a strong shift of maxima at high concentrations of antibodies (Ab) or their Fab fragment. This can be explained by a special type of Ab or Fab dimerization. Fluorescence decays of CP are measured at different concentrations of Ab and different pH values. The following deconvolution procedure based on the non-linear least squares method reveals a two- exponential character of the fluorescence decay. Data obtained by polarized fluorescence spectroscopy allows us to attribute the slow component (16 ns) to the free CP and the fast one (2.5 ns) to the CP-Ab complex. The obtained dependence of the ratio of amplitudes of the fast and slow components on the ratio of concentrations of CP and Ab at different pH values makes it possible to register separately the binding at different centers within the Ab and investigation of the effects of cooperation in the Ab molecule. The following experiments with Pt-coproporphyrin reveal the ability of Ab to reduce quenching of long-lived phosphorescence of the ligand.

Paper Details

Date Published: 1 February 1995
PDF: 7 pages
Proc. SPIE 2329, Optical and Imaging Techniques in Biomedicine, (1 February 1995); doi: 10.1117/12.200906
Show Author Affiliations
Oleg I. Lobanov, Institute of Biochemistry (Russia) and Univ. of Jyvaskyla (Finland)
Alexander P. Savitsky, Institute of Biochemistry (Russia)
Mary Nelen, Moscow State Univ. (Russia)
Marina V. Demcheva, Institute of Biochemistry (Russia)
Anatoly K. Yatsimirsky, Moscow State Univ. (Russia)
Jouko E. Korppi-Tommola, Univ. of Jyvaskyla (Finland)

Published in SPIE Proceedings Vol. 2329:
Optical and Imaging Techniques in Biomedicine
Hans-Jochen Foth; Aaron Lewis; Halina Podbielska; Michel Robert-Nicoud; Herbert Schneckenburger; Anthony J. Wilson, Editor(s)

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