Purification and biochemical characterisation of a novel breast carcinoma associated mucin-like glycoprotein defined by antibody 3E1.2

Stacker, Steven A.; Tjandra, Joe J.; Xing, Pei‐Xiang; Walker, Ian D.; Thompson, Christopher H.

doi:10.1038/bjc.1989.111

Cited by 13 publications

(4 citation statements)

References 39 publications

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“…A similar observation has been made for other glycoproteins with a small protein component [27,28]. (b) The size of the antigen is reduced by proteases with broad specificity (e.g.…”

Section: Discussionsupporting

confidence: 70%

Characterization of the CAMPATH‐1 (CDw52) antigen: biochemical analysis and cDNA cloning reveal an unusually small peptide backbone

et al. 1991

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The CAMPATH-1 (CDw52) antigen has been purified from human spleen. The antigenic epitope is heat stable but sensitive to mild alkali treatment. Experiments with phosphatidylinositol-specific phospholipase C indicate that it is anchored by a glycosylphosphatidylinositol (GPI) anchor. An N-terminal sequence of 11 amino acids was determined, followed by an abrupt stop. Using short overlapping mixed oligonucleotide primers, cDNA synthesized from the mRNA of a human B cell line was amplified by the polymerase chain reaction. The product was used to isolate cDNA clones and the full amino acid sequence of the CAMPATH-1 antigen was deduced. It consists of 37 amino acid residues plus a 24-residue signal peptide. It has all the features expected for a GPI-anchored membrane protein except that the predicted mature protein is remarkably short, comprising no more than 18 residues and possibly as few as 12 (depending on the GPI linkage site). Potential attachment sites for carbohydrate are present and it is shown that the antigen contains N-linked oligosaccharide(s). This structure accounts for the known properties of the antigen, though the exact reasons why it is such a good target for cell lysis in vitro and in vivo are not yet clear.

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“…A similar observation has been made for other glycoproteins with a small protein component [27,28]. (b) The size of the antigen is reduced by proteases with broad specificity (e.g.…”

Section: Discussionsupporting

confidence: 70%

Characterization of the CAMPATH‐1 (CDw52) antigen: biochemical analysis and cDNA cloning reveal an unusually small peptide backbone

et al. 1991

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“…bound. Ashall et al, 1982;Hilkens et al, 1984;Papsidero et al, 1984;Sekine et al, 1985;Price et al, 1986;Linsley et al, 1986;Stacker et al, 1989). cDNA clones coding for the core protein of this mucin, termed Polymorphic Epithelial Mucin (PEM), were recently isolated and the amino acid sequence was fully determined (Gendler et al, 1988 and.…”

Section: Discussionmentioning

confidence: 99%

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

Pancino

Osinaga²,

Charpin³

et al. 1991

Br J Cancer

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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.,

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“…While the 3E1 •2-defined epitope appears to be unique in that it is not expressed in components of human milk, MSA molecules are a member of the family of high molecular weight glycoproteins (1). The 3E1 -1 antibody was used to construct a serum assay which detected elevated antigen, mammary serum antigen (MSA), levels in 75% of patients with localized breast cancer and 90% of patients with metastatic breast cancer; only 2% of normal controls and 18% of patients with benign breast disease had elevated MSA levels (12.13).…”

Section: Introductionmentioning

confidence: 99%

Monoclonal antibodies reactive with mucin expressed in breast cancer

et al. 1989

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Summary Three murine monoclonal antibodies (BC 1, BC2 and BC3) were developed against human milk fat globule membrane (HMFGM). By immunoperoxidase staining, it was found that the antigenic determinants had a predominant distribution in breast cancer tissue. In addition, the antibodies reacted preferentially with mucin derived from human milk rather than that derived from the breast cancer cell line ZR75; they also recognized polymorphic high molecular weight components (MW>23OOOO) in serum and in human milk fat globule membrane. Thus the antibodies appear to react with a component of the family of mucins found in breast cancer and human milk and it appears likely that at least part of each epitope is protein in nature. Antibodies BCI, BC2 and BC3 recognized related but not identical epitopes, and they appear to be co-expressed on the same molecules as 3EI 2-defined antigen (mammary serum antigen, MSA) which is also a member of the family of breast cancer-rctated mucin. However, the 3E1 2 epitope is distinct and non-cross-reactive with those described for BCI, BC2and BC3. The BC2 and BC3 defined epitopes were examined for their value in serum assays. Immunoassay was developed with a combination of two antibodies, using antibody BC3 for antigen capture and antibody BC2 or 3E1 2 for antigen detection and gave reasonable sensitivity (-~85%) and specificity (~95%) in such serum tests for breast cancer. In a limited study, these tests appeared to complement the MSA test in the detection of breast cancer.

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Purification and biochemical characterisation of a novel breast carcinoma associated mucin-like glycoprotein defined by antibody 3E1.2

Abstract: Summary A member of the high molecular weight glycoproteins of human milk and breast cancer was isolated from the sera, ascites and breast carcinoma tissue of patients with breast cancer using monoclonal antibody 3E1.

Cited by 13 publications

References 39 publications

Characterization of the CAMPATH‐1 (CDw52) antigen: biochemical analysis and cDNA cloning reveal an unusually small peptide backbone

Characterization of the CAMPATH‐1 (CDw52) antigen: biochemical analysis and cDNA cloning reveal an unusually small peptide backbone

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

Monoclonal antibodies reactive with mucin expressed in breast cancer

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