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Chemiluminescence Quantitative Immunohistochemical Determination of MRP2 in Liver Biopsies

Massimo Guardigli, Marianna Marangi, Silvia Casanova, Walther F. Grigioni, Enrico Roda and Aldo Roda

Department of Pharmaceutical Sciences (MG,AR), Pathology Unit of the "F. Addarii" Institute of Oncology (MM,SC,WFG), and Department of Internal Medicine and Gastroenterology (ER), University of Bologna, Bologna, Italy

Correspondence to: Prof. Aldo Roda, Department of Pharmaceutical Sciences, University of Bologna, Via Belmeloro 6, I-40126 Bologna, Italy. E-mail: aldo.roda{at}unibo.it

Evaluation of protein expression in tissues and cells by electrophoretic and blotting techniques or by the quantification of the mRNA coding for the target protein is a common procedure in biochemistry research and clinical diagnoses. In this article, an alternative approach, based on an immunohistochemical procedure with chemiluminescent imaging detection, is described. The assay exploited the peculiar characteristics of the chemiluminescent detection of enzyme labels (high sensitivity and specificity, low background, easy quantification of the signal) for performing the direct, simple, and rapid quantitative evaluation of protein expression in tissues. When applied to the study of the levels of MRP2, a member of the human multidrug resistance–associated protein family, in samples obtained from formalin-fixed, paraffin-embedded liver biopsies, it allowed the reliable evaluation of the protein content of the tissue. Moreover, the analysis of clinical samples from patients with primary biliary cirrhosis under therapy with ursodeoxycholic acid gave results in line with those, previously reported in the literature, obtained with conventional protein expression analysis techniques. (J Histochem Cytochem 53:1451–1457, 2005)

Key Words: biopsy • chemiluminescence • MRP2 • imaging • immunohistochemistry • liver • protein expression

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