Quantitative Immunofluorescence Estimation of Pgp Expression in Human Solid Tumors by Flow Cytometry

摘要

An immunofluorescence assay that enables the quantitative detection of the level and intensity of the expression of multidrug resistance marker Pgp in human solid tumors 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 one to verify results, even after the tumor material has been stored for 2 years. The cost of analysis can be decreased by increasing the period of cell incubation with antibodies and reducing the antibody concentration. The possibility of analyzing minimal tumor samples, e.g., endoscopic biopsy specimens, by decreasing the concentration of cells incubated with antibodies to 100000 cells/mL and the number of cells used in the flow cytometer to 1000 has also been shown. Despite its continuance, the developed assay does not require any changes in the investigator's work schedule.