Carcinoma-Associated MUC1 Detected by Immunoradiometric Assays

Norum, Lars F.; Varaas, Tone; Kierulf, Bente; Nustad, Kjell

doi:10.1159/000056515

Cited by 14 publications

(12 citation statements)

References 24 publications

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“…This new automated assay will facilitate the rapid evaluation of new antibody combinations to MUC1 using individual patient sera rather than serum pools, and will be of considerable benefit in the analysis of the promising antibody combinations previously identified by the ISOBM MUC1 Workshop [13].…”

Section: Discussionmentioning

confidence: 99%

“…It was shown that some of these antibodies had specificity for carbohydrate epitopes, whereas the majority of the antibodies were directed to peptide epitopes. We tested all these antibodies in immunometric assays as both solid-phase and tracer antibodies using serum pools from cancer patients and healthy women [13]. A large number of antibody combinations were revealed as potential candidates for new promising 'cancer-specific' assays, which subsequently require testing on individual patient samples.…”

Section: Introductionmentioning

confidence: 99%

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Automated Immunofluorometric Assay for MUC1

Norum¹,

Nilsson²,

Nustad³

2001

Tumor Biol

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The aim of the present study was to establish a robust, reliable and fully automated immunofluorometric assay for the breast cancer serum marker MUC1. This would further serve as a prototype assay for evaluation of other MUC1 assays based on new antibody combinations. Using time-resolved fluorescence as tracer signal we developed an automated immunofluorometric assay for MUC1 (MUC1 IFMA). This assay was compared with two commercial assays. The CA15-3 EIA (CanAg) which use the same antibodies as the MUC1 IFMA, and the ETI-CA-15-3 K (Sorin) which use the original antibodies defining the CA 15-3 assay. The three assays showed comparable results. The coefficient of variation was below 10% from 9 to 2,400 kU/l for the MUC1 IFMA, from 15 to 250 kU/l for the CA15-3 EIA, and from 25 to 200 kU/l for the ETI-CA-15-3 K assay. At a specificity of 0.94 the overall diagnostic sensitivities for the MUC1-IFMA, CA15-3 EIA and ETI-CA-15-3 K assays were 0.40, 0.37, and 0.38, respectively. When applied to metastatic breast cancer, all assays had sensitivities close to 0.80. There was a close correlation (Spearman rank = 0.99) between results from the new assay and the CA15-3EIA. The new automated assay was not strictly immunometric as we could not achieve conditions where solid phase or tracer antibodies were in apparent excess. However, the assay performed well at a wide range of assay conditions. The automation, which minimizes imprecision in pipetting and handling of samples, and the high capacity of the AutoDELFIA instrument enabling measurement of all samples in a single run, were important aspects for establishing a reliable assay. The principle of the new automated immunofluorometric assay will be used as a rapid and reliable evaluation of a wide range of monoclonal antibody combinations in our search for the optimal MUC1 assay. This new automated immunofluorometric assay will be useful in the rapid and reliable evaluation of a wide range of monoclonal antibody combinations in our search for the optimal MUC1 assay.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Automated Immunofluorometric Assay for MUC1

Norum¹,

Nilsson²,

Nustad³

2001

Tumor Biol

Self Cite

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“…In a previous workshop of the International Society for Oncodevelopmental Biology and Medicine [15], more than 50 antibodies were tested both as solid-phase and tracer antibodies [16]. Fourteen of these antibodies were considered worthwhile for further testing in immunometric assays.…”

Section: Methodsmentioning

confidence: 99%

“…The International Society for Oncodevelopmental Biology and Medicine has previously organised a workshop for the characterisation of a large panel of antibodies against MUC1, with particular emphasis on the effects of altered glycosylation [15]. The panel of antibodies were also tested in immunometric assays as both solid phase and tracer antibodies, resulting in approximately 2,500 combinations [16]. Results from this work produced 104 antibody combinations that appeared to differentiate between MUC1 in two serum pools from cancer patients and normal controls.…”

Section: Introductionmentioning

confidence: 99%

New Immunoassays for MUC1 in Breast Cancer

Norum¹,

Sauren²,

Rye³

et al. 2001

Tumor Biol

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Eleven experimental immunofluorometric assays (IFMAs) were made using antibodies previously tested for epitope specificities. These assays were compared with six commercially available immunoassays. The clinical performance of these experimental assays was evaluated by analysing sera from 138 breast cancer patients and 105 female blood donors. The clinical performance of these assays was evaluated at a set specificity of 0.94. The highest overall sensitivity (0.56) was observed in the experimental assay with the antibody BC2 as solid phase and GP1.4 as the tracer antibody. This combination also showed the highest sensitivity in stage I/II breast cancer. The Truquant assay (Biomira) had an overall sensitivity of 0.51, and the highest sensitivity in stages III and IV at 0.65 and 0.94, respectively. The remaining commercial assays, with sensitivity ranging from 0.67 to 0.79, were below the top five experimental assays that showed sensitivity values between 0.79 and 0.85. The findings from our current study suggest that further development in MUC1 immunoassays could improve the detection of relapse in breast cancer patients.

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“…The mouse monoclonal antibody MB2 was raised against MCF7/AZ cells and selected for functional inactivation of E-cadherin. 26 Mouse monoclonal anti-MUC1 (214D4 27 ) and anti-MUC5AC (21M1 22 ) antibodies, and rabbit polyclonal anti-E-cadherin antibody (␣-UMT) were a gift, respectively, from Dr. J. Hilkens (The Netherlands Cancer Institute, Amsterdam), Dr. J. Bara (INSERM U-482, Saint-Antoine Hospital, Paris, France) and Dr. J. Behrens (Friedrich-Miescher Laboratory der Max Planck society in Tübingen). The polyclonal anti-MUC5AC antibody (LUM5-1 28 ) was a gift from Dr. Carlstedt (Lund University, Sweden).…”

Section: Antibodiesmentioning

confidence: 99%

Requirement of both mucins and proteoglycans in cell‐cell dissociation and invasiveness of colon carcinoma HT‐29 cells

Truant

Bruyneel

Gouyer

et al. 2003

Intl Journal of Cancer

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Key words: colon carcinoma cells; E-cadherin; mucins; heparan sulfate proteoglycansEpithelial E-cadherin, a component of the adherens junctions, is crucial for the organization and maintenance of differentiated epithelia. 1 The extracellular NH2-terminal domain of this transmembrane glycoprotein mediates cell-cell adhesion in a homophilic and Ca 2ϩ -dependent way. The COOH-terminal domain binds to the actin cytoskeleton via ␣-catenin and ␤-catenin or plakoglobin. 2 Studies in cancer cell lines have shown that changes in the structure or in the expression of the components of the E-cadherin/␤-catenin complex result in the suppression of cell-cell adhesion and in the expression of an invasive phenotype. [3][4][5][6][7][8] In cancer, the maintenance of the epithelia is lost and dissociation of cells is associated with metastatic dissemination. This phenomenon can occur through a decrease in the expression level of E-cadherin and, regarding colon cancer, such decrease has been related to the tumor grade and clinical outcome. 9 At the genomic level, loss of E-cadherin expression does not occur as a result of mutation of E-cadherin gene, whereas mutations of ␤-catenin were found in a small proportion of colon cancers. 10 -12 In another way, E-cadherin-mediated cell-cell adhesion may be modulated by the mucin-like glycoprotein MUC1, which is often overexpressed in cancer. MUC1, also termed episialin, is a membrane-associated glycoprotein composed of a membrane anchor and a mucin-like ectodomain. 13 In human ZR-75-1 breast cancer cells, the cytoplasmic domain of MUC1 was shown to bind ␤-catenin, leading to a decrease of E-cadherin-catenin complex and an anti-adhesion effect. 14 On the other hand, MUC1 was shown to cause steric hindrance of adhesion molecules like E-cadherin, through its large negatively charged extracellular domain, which contains many oligosaccharide side chains. 15,16 Proteoglycans were also shown to disturb the function of E-cadherin through steric hindrance in variants of the murine mammary gland cell line NMuMG and the canine epithelial kidney cell line MDCK. 17 During colorectal carcinogenesis, the expression of MUC1 was found upregulated, particularly in later stages of the disease. 18 -20 On the other hand, de novo appearance of a secreted mucin normally found in the stomach, MUC5AC, has been described in colon carcinomas. [21][22][23] Thus, mucinous differentiated cells are present in colorectal carcinomas and more particularly in mucinous carcinomas and in signet ring cell carcinomas. Until now, the significance of a differentiation characteristic into a mucin-secreting phenotype with regard to the tumor development is still unknown.To investigate the role of mucins in the invasive properties of colon carcinoma cells, the HT-29 cell line is an appropriate model, as the cells of this cell line express different phenotypes: besides a majority of undifferentiated cells (Ͼ95%), a small proportion of cells presents a capacity to differentiate into a mucinous phenotype or into an enterocyte-like ...

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Carcinoma-Associated MUC1 Detected by Immunoradiometric Assays

Cited by 14 publications

References 24 publications

Automated Immunofluorometric Assay for MUC1

Automated Immunofluorometric Assay for MUC1

New Immunoassays for MUC1 in Breast Cancer

Requirement of both mucins and proteoglycans in cell‐cell dissociation and invasiveness of colon carcinoma HT‐29 cells

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