Validation of a simple avidin-biotin detection method for Helix pomatia lectin (HPA) binding as a prognostic marker in cancer

Abstract

Helix pomatia lectin (HPA) binding is a marker of metastatic competence in several human cancers. The altered cellular glycosylation detected by HPA is of clinical interest and functional significance, but research has been hampered by uncertainty over appropriate and accessible histochemical methods. Most studies have employed a complex multi-layered detection system localising binding of unconjugated HPA by layering with a polyclonal antibody to HPA, a biotinylated secondary antibody against the first antibody and streptavidin peroxidase. This detection system is sensitive and yields accurate prognostic information, but is lengthy and requires antibodies against HPA that are not widely available. A simpler technique, that uses peroxidase-labelled HPA is inappropriate as the carbohydrate-combining characteristics of the lectin are altered, and the prognostic significance of lectin binding is lost. Therefore a valid alternative, simple and accessible technique is required. In the present study, we compare the results of labelling of HPA binding using the complex multi-layered detection system with a simple avidin-biotin method. In a series of 101 breast cancers, both methods gave comparable results. Therefore, the avidin-biotin method appears to be appropriate for studies on HPA binding to detect altered glycoforms in cancer. It is hoped that its adoption may encourage research into this clinically significant alteration in cellular glycosylation.