Epithelial membrane antigen: Partial purification, assay and properties

Ormerod, M.G.; Steele, Kate; Westwood, J. H.; Mazzini, María N.

doi:10.1038/bjc.1983.226

Cited by 86 publications

(25 citation statements)

References 18 publications

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“…Full characterization of the epitopes bound by M3 and M24 has however proved difficult but it seems that neither of these antibodies reacts with the Y-or the Forsmann antigen, red blood cells from groups A, B or 0, or with red blood cells carrying the Lewis antigens Lewisa+b-, Lewisa-b+, Lewisa-b-(R. A. J. McIlhinney, unpublished work). The epitope bound by M3 and M24 also do not seem to be that recognized by the monoclonal antibody SSEA-1, Galfl1-4(Fucal-3)GlcNAcfll-6 (Gooi et al, 1981) (Ormerod et al, 1983). These results also suggest that M8 binds to a complex epitope involving a short sequence of amino acids and sugars (M. G. Ormerod, J. H. Westwood & K. Steele, personal communication).…”

Section: Discussionsupporting

confidence: 53%

Monoclonal antibodies recognizing epitopes carried on both glycolipids and glycoproteins of the human milk fat globule membrane

McIlhinney¹,

Patel²,

Gore³

1985

Biochemical Journal

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The molecules of the human milk fat globule membrane (MFGM) which bind four murine monoclonal antibodies (LICR LON M3, M8, Ml 8 and M24) raised against the human MFGM have been identified. By using 'Western' blotting [Burnette (1981) Anal. Biochem. 112,[195][196][197][198][199][200][201][202][203] it was shown that each antibody reacted with a different set of proteins. M3 and M24 were similar in their pattern of reaction with the membrane proteins, but were quite distinct from M8 and M18, which also differed from each other. Glycopeptides prepared from the MFGM by exhaustive Pronase digestion were able to inhibit partially the binding of M3 and M24, and prevent totally the binding of M8 and M18, to the MFGM in an enzyme-linked immunoabsorbent assay. Oligosaccharides obtained by the deproteination of human milk also completely inhibited the binding of M3, M18 and M24 to the MFGM. However, the binding of M8 was not inhibited by these saccharides, and therefore M8 may not be recognizing a simple carbohydrate determinant. By using an enzymelinked assay, M8 and M18 were shown not to bind to MFGM glycGlipid, whereas M3 and M24 did, and this was confirmed by overlaying thin layer chromatograms of MFGM lipids with these antibodies. Both M3 and M24 showed a similar complex pattern of reaction, binding to more than one glycolipid moiety. By these means all four antibodies have been shown to react with antigens which involve carbohydrate side chains carried on different proteins, and two were also shown to react with such determinants on glycolipids.

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Section: Discussionsupporting

confidence: 53%

Monoclonal antibodies recognizing epitopes carried on both glycolipids and glycoproteins of the human milk fat globule membrane

McIlhinney¹,

Patel²,

Gore³

1985

Biochemical Journal

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“…To determine the amount in the patients sera a competitive radioimmunoassay was used that is based on the binding of '25I-ICR2 to plates coated with purified EMA (Ormerod et al, 1983). Serial dilutions of sera in phosphate buffered saline containing 0.5% bovine serum albumin (PBS/ BSA) were made in triplicate and 30 JAl aliquots of each were mixed with an equal volume of '25I-ICR2 (105 c.p.m.…”

Section: Radiolabelled Antibodymentioning

confidence: 99%

The effect of circulating antigen and radiolabel stability on the biodistribution of an indium labelled antibody

Davidson

Babich²,

Young³

et al. 1991

Br J Cancer

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Summary This study has investigated two of the main problems with radiolabelled antibody imaging, the formation of circulating immune complexes (I.C.) and the non specific binding of radiolabel to the antibody molecule.Patients undergoing immunoscintigraphy with "'In labelled monoclonal antibody ICR2 were divided into three groups who received either the radiolabelled antibody alone (control, n = 12), the radiolabelled antibody which was incubated with the chelating agent diethylene triamine pentacetic acid (DTPA) prior to size exclusion chromatography (n = 6) or whose injectate was treated with DTPA and cold MAb administered intravenously prior to radiolabelled MAb administration (n = 6). Radiolabelled antibody uptake in abdominal organs was measured by region of interest analysis using a gamma camera with online computer and that in tumour and normal tissues by gamma well counting of biopsies. Circulating antigen and immune complex was measured by high pressure liquid chromatography (HPLC).The sensitivity of tumour imaging and the tumour uptake of radiolabelled antibody was not significantly different between the groups. Patients with high circulating antigen levels developed high levels of circulating immune complex but also had high tumour uptakes of radiolabelled antibody. Administration of cold MAb increased the splenic, but did not effect the tumour uptake of radiolabelled antibody and only minimally reduced levels of circulating immune complex. Chelate administration reduced the urinary excretion of radioactivity but increased the liver uptake of radioactivity.These results have shown that successful antibody imaging can be carried out despite high levels of circulating antigen, that large doses of unlabelled antibody are required to prevent immune complex formation and that removal of non specifically bound "'In does not reduce the liver uptake of radioactivity.

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“…A number of monoclonal antibodies have been described which react with breast cancer associated antigens and have tissue distribution similar to 3E1.2 Ellis et al, 1984;Hilkens et al, 1984;Kufe et al, 1984;Papsidero et al, 1983). Most of these antibodies have been shown to detect circulating antigens in the serum of patients with advanced breast cancer but the detection in localised breast cancer has been poor (Burchell et al, 1984;Dhokia et al, 1986;Hayes et al, 1985;Hilkens et al, 1984Hilkens et al, , 1986Kufe et al, 1984).…”

mentioning

confidence: 99%

“…PAS-0 component has also been referred to as EMA (epithelial membrane antigen) complex (Ormerod et al, 1983(Ormerod et al, , 1985 and recently popularised as human PEM (polymorphic epithelial mucin) because of the genetic polymorphism exhibited.…”

mentioning

confidence: 99%

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

Stacker

Tjandra²,

Xing³

et al. 1989

Br J Cancer

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Epithelial membrane antigen: Partial purification, assay and properties

Cited by 86 publications

References 18 publications

Monoclonal antibodies recognizing epitopes carried on both glycolipids and glycoproteins of the human milk fat globule membrane

Monoclonal antibodies recognizing epitopes carried on both glycolipids and glycoproteins of the human milk fat globule membrane

The effect of circulating antigen and radiolabel stability on the biodistribution of an indium labelled antibody

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

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