Characterization of the shed form of the human tumor-associated glycoprotein (TAG-72) from serous effusions of patients with different types of carcinomas
| Autor: | R S, Katari, P D, Fernsten, J, Schlom |
|---|---|
| Rok vydání: | 1990 |
| Předmět: |
Rectal Neoplasms
Transplantation Heterologous Radioimmunoassay Antibodies Monoclonal Ascites Mice Nude Enzyme-Linked Immunosorbent Assay Antigen-Antibody Complex Exudates and Transudates Cell Line Pancreatic Neoplasms Endometrium Mice Antigens Neoplasm Colonic Neoplasms Chromatography Gel Animals Humans Female Neoplasm Transplantation Glycoproteins |
| Zdroj: | Cancer research. 50(16) |
| ISSN: | 0008-5472 |
| Popis: | Monoclonal antibody (MAb) B72.3 binds a high molecular weight tumor-associated glycoprotein designated TAG-72. This study reports the isolation and characterization of secreted TAG-72 directly from effusions of ovarian, colorectal, pancreatic, and endometrial carcinoma patients and compares them to TAG-72 derived from the LS-174T colon carcinoma xenograft. The B72.3-reactive antigen, TAG-72, was used as immunogen to produce second generation anti-TAG-72 MAbs. One of these second generation MAbs, CC49, had a higher affinity than that of B72.3 and was utilized as an affinity reagent in a procedure to purify the TAG-72 present in the serous effusions of carcinoma patients. A three-step purification procedure, utilizing heat extraction, CC49 antibody affinity chromatography, and gel filtration chromatography, resulted in 1000- to 4400-fold purifications of the TAG-72 derived from effusions, as analyzed using a double-determinant radioimmunoassay. Radiolabeled TAG-72 from each of the effusions demonstrated similar high molecular weight bands on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Similar results from the various effusions were also obtained in Western blotting analyses. Analyses of TAG-72 from the different effusions in radioimmunoassay using five different anti-TAG-72 MAbs revealed similar binding patterns. The results of these studies thus indicate that TAG-72 obtained directly from patients with ovarian, colorectal, endometrial, and pancreatic carcinomas and the LS-174T xenograft are highly similar in terms of immunochemical properties and antigenic profile. |
| Databáze: | OpenAIRE |
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