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Purification and immunochemical characterization of ascitic fluid glycoproteins containing certain tumor-associated and blood group antigen markers

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Abstract

Ascitic fluids from patients with various types of cancer were screened for the CA 19-9 and CA 125 tumor-associated antigenic activities. Two fluids exhibiting the highest activities were tested for their binding to various lectin-Sepharose columns resulting in both being bound best to wheat germ agglutinin (WGA) Sepharose. The WGA column eluate of one fluid was further chromatographed by HPLC and three peaks were obtained with approximate molecular weights of 3.65 MDa, 664 kDa and 330 kDa, of which only the largest fraction contained the CA 19-9 activity. The fluids were also fractionated on a Sephacryl S-400 column with most of the activity being present in or near the void volume.

Monoclonal antibodies were used to demonstrate that the purified glycoproteins also contained the blood group A determinant, the four Lewis determinants Lea, Leb, Lex and Ley, and the sialylated-Lex determinant, while other antibody analyses failed to detect other blood group and/or carbohydrate sequence determinants. Some of the blood group expressions could be separated from the CA 19-9 and CA 125 active glycoproteins by adsorption with various lectins other than the WGA.

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Abbreviations

NeuAc:

N-acetyl-D-neuraminic acid

Gal:

galactose,D-galactopyranose

Fuc:

fucose,L-fucopyranose

GlcNAc:

N-acetyl-D-glucosamine

GalNAc:

N-acetyl-D-galactosamine

WGA:

wheat germ agglutinin

PBS:

phosphate buffered saline

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Authors and Affiliations

  1. Departments of Molecular Biology, and Otolaryngology and Head and Neck Surgery, Northwestern University Medical School, 60611, Chicago, IL, U.S.A.

    Mario Venegas, Loretta Liu, Laura Lovell, Lyman E Davis & Byron Anderson

  2. Cancer Research Laboratory, Abbott Laboratories, 60064, North Chicago, IL, U.S.A.

    Teresa Wilbanks, Michael Hass, George Manderino & Harry Rittenhouse

Authors
  1. Mario Venegas
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  2. Loretta Liu
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  3. Laura Lovell
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  4. Lyman E Davis
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  5. Byron Anderson
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  6. Teresa Wilbanks
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  7. Michael Hass
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  8. George Manderino
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  9. Harry Rittenhouse
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Venegas, M., Liu, L., Lovell, L. et al. Purification and immunochemical characterization of ascitic fluid glycoproteins containing certain tumor-associated and blood group antigen markers. Glycoconjugate J 6, 511–524 (1989). https://doi.org/10.1007/BF01053774

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  • DOI: https://doi.org/10.1007/BF01053774

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