Purification of the 67-kDa lectin-like glycoprotein of Trypanosoma cruzi (LLGP-67) based on its binding to non-derivatised agarose. Evaluation of the obtained protein as a relevant antigen for diagnosis of the infection

IVÁN S. MARCIPAR (EJECUTOR PRINCIPAL), ELINA WELCHEN, CINTIA ROODVELDT, ALBERTO J. MARCIPAR AND ARIEL M. SILBER

FEMS MICROBIOLOGY LETTERS

Año: 2003 vol. 220 p. 149 - 154

0378-1097

The setting up of affinity chromatography methods for one-step purification of antigens able to be used in diagnosis of infectious diseases has an intrinsec interest. We propose a simple direct method to affinity purify to aparent homogeneity a previously described antigen relevant for the diagnosis of infection by Trypanosoma cruzi, the aethiological agent of Chagas disease. This antigen consists of a 67 kDa lectin like glycoprotein (LLGP-67) that presents galactose binding activity. The purification strategy proposed in this report takes advantantage of the galactose binding activity of this protein, and the fact that non-derivatised agarose is a polymer containing exposed galactose residues. We describe results showing that this antigen can be purified from Trypanosoma cruzi total homogenates in a direct way using non-derivatised agarose as chromatographic ligand and demonstrate that the antigenic properties for human diagnosis of Chagasic infection of the agarose purified  protein is identical to that of LLGP-67 purified by other methods.