Specificity and sensitivity of immunocytochemistry for detecting P-glycoprotein in haematological malignancies.
OBJECTIVE
To determine the optimal working conditions of the alkaline phosphatase-antialkaline phosphatase (APAAP) method to establish a specific and sensitive assay for the detection of low numbers of MDR positive cells in patients with hematological malignancies.


METHODS
Three monoclonal antibodies (C-219, JSB-1, MRK-16) were used for the detection of P-glycoprotein (P-gp) in cell lines and in samples from 43 patients with haematological malignancies. The results of the APAAP method were compared with western blotting for specificity and sensitivity.


RESULTS
Excellent correlation was obtained between optimised APAAP and western blotting, except in the case of multiple myeloma. JSB-1 seemed to be the more useful monoclonal antibody for the APAAP which was more sensitive than western blotting in its ability to detect single P-gp positive cells.


CONCLUSIONS
Methods for P-gp detection, as defined by multidrug resistant (MDR) cell lines, are not necessarily optimal and specific for clinical samples and may lead to higher false positive and negative results, according to the conditions and the monoclonal antibodies used.
Gala
J L
JL
Haematology Department, Royal North Shore Hospital, Sydney, Australia.
McLachlan
J M
JM
Bell
D R
DR
Michaux
J L
JL
Ma
D D
DD
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D015153:Blotting, Western; D002352:Carrier Proteins / Q000032:analysis*; D006801:Humans; D007124:Immunoenzyme Techniques; D007945:Leukemia, Lymphoid; D007951:Leukemia, Myeloid; D008223:Lymphoma / Q000737:chemistry*; D008228:Lymphoma, Non-Hodgkin / Q000737:chemistry; D008562:Membrane Glycoproteins / Q000032:analysis*; D009101:Multiple Myeloma; D009363:Neoplasm Proteins / Q000032:analysis*; D012680:Sensitivity and Specificity; D014407:Tumor Cells, Cultured
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D002352:Carrier Proteins; D008562:Membrane Glycoproteins; D009363:Neoplasm Proteins