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T. A. Bogush, M. V. Tikchomirov, E. A. Dudko, M. N. Sinitsyna, R. J. Ramanauskaite, B. E. Polotsky, S. A. Tyulyandin, M. I. Davydov
Quantitative
immunofluorescence estimation of Pgp expression in human solid tumors by flow
cytometry
Abstract
An immunofluorescence assay that enables quantitative detection of
level and intensity of multidrug resistance marker Pgp expression in human
solid tumors by using flow cytometry and monoclonal antibody (clone 17F9, BD
Pharmingen) was developed and adapted for clinical assessment. The analysis
includes the following relevant task-specific stages. Single-cell suspension
can be prepared from both surgical biopsy native tumor tissues and tumor
specimens fixed in 4% formaldehyde. The assay allows to result verification
even after two years of tumor material storage. Cost of analysis can be
decreased by increasing of period of cell incubation with antibody and reducing
of the antibody concentration. It was shown also a possibility to analyze
minimal tumor samples, e.g. endoscopic biopsy specimens, by decreasing
concentration of cells incubated with antibodies to 1x105 cells/ml,
and flow cytometer counted cell number to 1x103. Although its
continuance, the developed assay does not require any changes in investigator
work schedule.
Copyright (C) Chemistry Dept., Moscow State University, 2002
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