HUMAN CARCINOMA-ASSOCIATED ANTIGEN (HCA), ISOLATED FROM THE ENDOMETRIAL CARCINOMA CELL-LINE KLE-1 AND ASCITIC FLUID OF A PATIENT WITH OVARIAN-CARCINOMA - COMPARISON WITH EPIGLYCANIN

Human carcinoma-associated antigen (HCA), detected by the mouse monocl onal anti-epiglycanin antibody, AE-3, has been isolated from ascitic f luid taken from a patient with metastatic ovarian adenocarcinoma and f rom spent medium of the human endometrial carcinoma cell line KLE-1 an d compared with epiglycanin. The ascitic fluid and the spent medium we re concentrated by a Filtron Ultrasette 100 K Omega membrane and fract ionated by gel filtration on Sepharose CL-2B. The active fractions whi ch consisted mainly of glycoproteins having relative molecular weights in the range 1000-2000 kDa compared to dextrans, were further purifie d by affinity chromatography on a column of immobilized AE-3. The acti ve fraction was subjected to SDS-PAGE and blotted onto a PVDF membrane . The amino acid composition of HCA isolated from the two sources, wer e related but not identical and both showed some differences from the amino acid composition of epiglycanin. They all have, however, composi tions typical of mucin-type glycoproteins. The isoelectric point for H CA from both KLE-1 and ascitic fluid were determined to be at pH 1.8 a nd the buoyant densities were about 1.4 g/ml as determined by cesium t rifluoroacetate gradient centrifugation. (C) 1998 Elsevier Science B.V .