Purification and characterisation of a breast-cancer-associated glycoprotein not expressed in normal breast and identified by monoclonal antibody 83D4

Abstract

The purpose of the present study was to purify and characterise the antigen identified by mAb 83D4. The antigen was isolated from different sources, and purified antigen was analysed by biochemical and immunological methods. Moreover, the reactivity of mAb HMFG-1 (TaylorPapadimitriou et al., 1981) defining the PEM antigen and of mAb B72.3 and CC49 (Muraro et al., 1988), defining the TAG-72 antigen, with the 83D4 purified antigen was investigated. Materials and methods mAbIgM mAb 83D4 was generated by immunisation of Balb/c mice with cell suspensions from formalin-fixed paraffinembedded sections of an invasive human breast carcinoma as described in detail elsewhere (Pancino et al., 1990a). The antibody was purified from ascitic fluid by dialysis against demineralised water (Garcia-Gonzales et al., 1988). Precipitated antibody was resuspended in 0.1 M Tris-HCI pH 8, 1 M NaCl and dialysed against the appropriate buffer.Control antibodies produced in our laboratory were BIN, an IgM reactive with a nuclear protein; CA4, an IgM raised to a human milk cell glycoprotein (Pancino et al., 1991) and 1BE12, an IgM mAb against a breast-cancer-associated glycoprotein (Pancino et al., 1990b). Culture supernatant containing mAb HMFG-1 (Taylor-Papadimitriou et al.,

Clinical Application of Serum Tumor Associated Material (TAM) from Non-small Cell Lung Cancer Patients

“…This finding was later corroborated with histological evidence that lectins show differential binding to healthy compared with malignant tissue (Turner, 1992;Saussez et al, 1998). More recently, cancer-associated cell-surface glycans have been directly characterized using specific monoclonal antibodies and mass spectrometry (Shriver et al, 2004), further illuminating the molecular changes that occur upon malignant transformation (Matsushita et al, 1990;Pancino et al, 1991).…”

Combination effect on HIV infection in vitro of soluble CD4 and HIV-neutralizing antibodies

“…As the natural antibodies in vivo display a heterogeneous mixture of various specificities, we have examined the effect of neutralizing anti-V3 antibodies (mouse IgG1 NEA-9205 from Dupont) in combination with sCD4 (Neosystem Laboratoire, Strasbourg) and antibodies which bind to the CD4-binding domain of gpl20 (human IgG1 MAb 1B1 from Waldheim Pharmazeutika, Vienna) or to other epitopes on gpl20. These latter antibodies were the broadly neutralizing human IgGIMAb 2G12, which binds to a carbohydrate dependent epitope on gpl20 but does not block CD4 binding (Waldheim Pharmazeutika), and a monoclonal mouse IgM MAb 83D4 raised against a human breast cancer mucin and binding to GalNAc-Ser/Thr [9,10], which has previously been described as a neutralization epitope of a broad range of HIV-1 strains I-5]. Additionally, we also tested a polyclonal heat-inactivated serum against gpl20…”

Carbohydrate-based vaccines with a glycolipid adjuvant for breast cancer

“…The aberrations include loss or overexpression of certain glycan structures, persistence of truncated glycans, and emergence of novel glycans. Some of the aberrant glycans on malignant tissues were identified by immunohistochemical staining with lectins (2,3) or mAb or by MS (4,5). To date, numerous tumor-associated antigens expressed on cancer cells in the form of glycolipids or glycoproteins have been characterized and correlated to specific types of cancers (6,7).…”