Differential expression of alpha2,6-sialyltransferase in colon tumors recognized by a monoclonal antibody.
Gangopadhyay A. Perera SP. Thomas P.
Laboratory of Cancer Biology, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, MA 02215, USA.
It has been reported that alpha2,6-sialyltransferase (alpha2,6-ST; E.C. 188.8.131.52) activity is associated with cellular differentiation. To define its role in colon carcinoma differentiation, we have generated murine monoclonal antibodies (MAb) against alpha2,6-sialyltransferase. The MAb, designated 6B9 of IgM isotype, showed strong reactivity with the purified and crude alpha2,6-ST by ELISA and dot blot assays. Western blotting with MAb 6B9 identified purified alpha2,6-ST of MW 47 kDa and the same MW protein from rat and human liver extracts. The MAb also reacted with two other liver proteins of approximate MW 65 and 100 kDa. Immunoperoxidase studies with formalin-fixed paraffin-embedded tissues showed that MAb 6B9 reacts with liver tissues, the staining of hepatocytes was granular and cytoplasmic. There was a distinct pattern of zonal distribution of this enzyme in hepatocytes located particularly in the portal areas of the liver corresponding to zone 1. Normal colon (100%) and hyperplastic polyps (100%) showed very weak to no reactivity. Adenomas (100%) demonstrated moderate reactivity, while the poor (33%), moderate (100%) and well-differentiated (80%) colon adenocarcinomas showed strong reactivity. Results suggest that alpha2,6-ST is associated with the differentiation state of colon tumors.
A monoclonal antibody against the X protein of hepatitis B virus: fine mapping of its epitope and application in a quantitative ELISA of the X protein in sera of hepatitis B patients.
Kumar V. Jayasuryan N. Reddi H. Sahal D. Panda SK.
Virology Group, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi, India.
A HBx-specific mouse monoclonal antibody was developed and its epitope mapped to a hydrophilic segment 94HKRTLGL100 using the multipin peptide synthesis technique. A sensitive ELISA with a threshold of 5 to 10 ng was developed to identify the HBx-positive hepatitis B cases and measure the levels of HBx in sera. The same patient sera were also analyzed for the presence of anti-HBx using the purified recombinant antigen. HBx was present in 23% of the cases (15/65) whereas only 14% of the cases (9/65) were positive for anti-HBx. The mean value of HBx in acute hepatitis sera was higher (522 ng/ml) than in cirrhosis cases (48 ng/ml). PCR amplification of the S gene showed that all 15 HBx-positive cases were also positive for the viral DNA.