Revista Colombiana de Química
versión impresa ISSN 0120-2804
VEGA, Nohora; MURCIA, Hansen y PEREZ, Gerardo. CHARACTERIZATION OF Salvia bogotensis ANTI-LECTINIGYS AND THEIR APPLICATION IN IMMUNOCYTOCHEMICAL STUDIES INVOLVING TN ANTIGEN DETECTION. Rev.Colomb.Quim. [online]. 2009, vol.38, n.3, pp. 363-377. ISSN 0120-2804.
Immunoglobulins isolated from egg yolk (IgY) are tools which are currently used in different fields of the biological sciences; they have clear advantages over mammalian serum IgGs. We have established the conditions for obtaining anti-Salvia bogotensis lectin IgYs in previous work; their use in immunocytochemical studies requires that their main molecular characteristics are known as well as the conditions for IgY-lectin interaction. Salvia bogotensis lectin (SBoL) can specifically recognise Tn antigen, a recognised tumoral marker in many types of cancer but additional tools are required for evidencing this interaction in cells. Given the availability of S. bogotensis anti-lectin IgY, this work was aimed at molecularly characterising these IgYs and evaluating their application in immunocytochemical studies for detecting Tn antigen in tumour cells. Purified IgYs' isoelectric points, molecular weight and carbohydrate content were determined. Homologous and heterologous lectins were obtained for establishing the specificity of antibody interaction with the lectin; they were assayed by ELLSA. Biotin-or peroxidase-labelled IgYs were prepared; Tn antigen was specifically detected by CELISA and immunocytochemistry in MCF-7 and HeLa cell-lines with the lectin which was revealed with the labelled IgYs. Results showed that anti-SBoL IgY antibodies represent a highly sensitive tool for specific Tn antigen recognition assays.
Palabras llave : Lamiaceae; Salvia bogotensis; lectin; IgY; Tn antigen; immunocytochemistry.