Reactivity of an anti-(human gastric carcinoma) monoclonal antibody with core-related peptides of gastrointestinal mucin

Price, Michael R.; Sekowski, M.; Yang, Gangyi; Durrant, Lindy G.; Robins, R. A.; Baldwin, Robert

doi:10.1007/bf01742533

Cited by 14 publications

(9 citation statements)

References 16 publications

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“…As well as overexpression of MUC1, altered glycosylation of the VNTR is responsible for the exposure of these peptide epitopes in tumours (Gendler et al, 1988;Devine et al, 1990a). *Despite the success of MUC1 VNTR-reactive MAbs, only a few MAbs reacting with other VNTR have been reported Price et al, 1991), and the use of these MAbs for diagnosis and therapy has not been investigated. The MUC2 mucin is of particular interest since this is a major component of mucus produced by patients with colon and lung cancer, as well as those with cystic fibrosis (Gum et al, 1989;Gerard et al, 1990;Jany et al, 1991).…”

mentioning

confidence: 99%

Monoclonal antibodies reacting with the MUC2 mucin core protein

Pl¹,

McGuckin²,

Birrell³

et al. 1993

Br J Cancer

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Summary This study sought to produce monoclonal antibodies (MAbs) which reacted with the MUC2 core protein. Two MAbs [3A2 (IgGl) and 4F1 (IgM)] were produced by immunising female BALB/c mice with gel-formed mucin from the LS174T colon cancer cell line followed by a KLH conjugate of a 29 amino acid synthetic peptide whose sequence was derived from the variable number of tandem repeats (VNTR) region of a MUC2 cDNA clone.The MAbs reacted with synthetic MUC2 VNTR peptides but not synthetic MUCI or MUC3 VNTR peptides, and showed specific reactivity in Western blotting with a high molecular weight protein produced by the LS174T colon carcinoma cell line. The use of shorter peptides indicated that the minimum peptide epitopes for these MAbs were different. Mab 3A2 reacted with amino acids 5-19 of the MUC2 VNTR by inhibition ELISA but not by direct ELISA, while 4FI reacted with this peptide in both assays. Furthermore, 4FI reacted in direct ELISA when a larger (29 amino acid) MUC2-derived peptide was coated onto the assay plate by incubating in carbonate buffer or by drying the peptide onto the assay plate, while 3A2 only reacted when this peptide was coated in carbonate buffer. The different specificity of the MAbs was also illustrated by the reactivity of 4F1 but not 3A2 with partially deglycosylated cystic fibrosis mucin.Immunohistochemical analysis with these MAbs revealed a strong reactivity with lung, gastric and colon tumours relative to normal tissue, with some breast and ovarian tumours also reacting. Both MAbs stained some normal goblet cells in the perinuclear region but not the mucin droplet or secreted mucin, indicating a reaction with immature (poorly glycosylated) mucin in the endoplasmic reticulum and/or golgi, but not with mature (fully glycosylated) mucin. In contrast, tumours showed strong diffuse cytoplasmic staining. 4F1 also showed weak apical cytoplasmic staining in some goblet cells and stained some tumours which showed no reactivity with 3A2.These antibodies should prove useful in the study of MUC2 structure and function, and in the diagnosis of some tumours.

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mentioning

confidence: 99%

Monoclonal antibodies reacting with the MUC2 mucin core protein

Pl¹,

McGuckin²,

Birrell³

et al. 1993

Br J Cancer

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“…It was generated by the immunization of BALB/c mice with dissociated gastric carcinoma cells isolated from a lymph node metastasis of a well-differentiated adenocarcinoma (39). This antibody was later found to react with the epitope TTTTT within the tandem repeat region of MUC2 (40). Subsequently, purified chemically deglycosylated colon cancer mucins were utilized by Gambus et al (41) to generate MUC2 antibodies.…”

Section: Muc2mentioning

confidence: 99%

Mucin antibodies - new tools in diagnosis and therapy of cancer

et al. 2001

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Many cancer and diseased cells are distinguished from their normal counterparts by an altered expression of cell-surface epitopes. One family of molecules that show altered expression on tumor cells is mucins (MUC). Unlike normal tissue where MUC exists as heavily glycosylated form, the disease- or tumor-associated MUC molecules are underglycosylated. Such underglycosylation of the core protein in cancer tissues exposes new epitopes on the cell surface that are unique to cancer tissues. Several monoclonal antibodies (Mabs) have been generated against these normal and tumor-associated mucins. Enzymatic fragments of Mabs like F(ab')2 and Fab have shown improved clinical utility for diagnosis, imaging, and therapy of cancer. Genetic-engineering methods have been used to design antibody fragments exhibiting high functional affinity, good tumor localization, and rapid clearance from the blood stream thus minimizing radiation exposure to the normal tissues. Such recombinant fragments have shown encouraging results in preclinical studies using xenografted tumor bearing mice and present a whole new avenue for radioimmunotherapy and diagnosis of cancer.

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“…Gastrointestinal cancer is a common cause of death worldwide [1]. In the absence of metastasis most cancers would be cured by surgery, chemotherapy disease [4] and might enable a renaissance in clinical diagnosis [2].…”

Section: Introductionmentioning

confidence: 99%

“…The protein core consists of a variable number of tandem repeats with the consensus amino acid sequence of 1 PTTTPITTTTTVTPTPTPTGTQT 23 [5]. Previously several monoclonal antibodies (mAbs) have been developed in mice using a synthetic peptide (K 12 VTPTPTPTGTQTPT 25 ) derived from the repeat region of MUC2 and conjugated to KLH (keyhole limpet haemocyanin) carrier [4]. Some of these antibodies (such as mAb 994 and 996, both belonging to the IgG1 subclass of antibodies) have recognized samples from human tumour tissues of the colon, as well as soluble tumour derived immunoaffinity purified MUC2 mucin [7].…”

Section: Introductionmentioning

confidence: 99%

Heteroclitic recognition of combinatorial TX¹TX²T peptide mixtures by mucin‐2 protein specific monoclonal antibody

Windberg¹,

Uray²,

Illyés³

et al. 2003

Journal of Peptide Science

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The mucin-2 (MUC2) glycoprotein secreted by the epithelial cells of human colon may be abnormally under-glycosylated in the case of cancer. Monoclonal antibody (mAb) 994 raised against the immunogenic part of the protein core, recognizes malignant human colon tissues as well as pentapeptides with TX1TX2T motif present in MUC2. Using a combinatorial approach and ELISA experiments it was found that mAb 994 is able to recognize peptides of the sub-library TQTX2T very strongly, and to some extent also peptides from TETX2T, TLTX2T and TVTX2T sub-libraries. Binding studies with peptides corresponding to the TQTX2T and TETX2T sub-libraries showed that mAb 994 recognized only six peptides (IC50 = 9-208 micromol dm(-3)) from the 19 compounds of the TQTX2T sub-library and only three peptides (IC50 = 3500-16700 micromol dm(-3)) from the 'second-best' TETX2T sub-library. The most pronounced mAb binding occurred when Gln was in position X1 and it was much weaker in the case of Glu, Val or Leu. As for X2 amino acids, the presence of Pro, Ala can provide a strong, while Tyr, Trp, Phe and Ser a weaker, peptide-antibody interaction. Data from this study suggest that pentapeptide TQTPT, whose sequence is present in the native protein, is bound most strongly. However, almost identical binding properties were observed with peptide TQTAT, whose sequence is not present in the protein. Apart from this, some other 'heteroclitic' peptides were found with a different rank in the binding-hierarchy. Based on these peptides artificial compounds can be prepared as potential candidates for vaccine development. Results of this study also provide a rationale for understanding the molecular background of the heteroclitic nature of the MUC2 protein core specific mAb 994.

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Reactivity of an anti-(human gastric carcinoma) monoclonal antibody with core-related peptides of gastrointestinal mucin

Cited by 14 publications

References 16 publications

Monoclonal antibodies reacting with the MUC2 mucin core protein

Monoclonal antibodies reacting with the MUC2 mucin core protein

Mucin antibodies - new tools in diagnosis and therapy of cancer

Heteroclitic recognition of combinatorial TX¹TX²T peptide mixtures by mucin‐2 protein specific monoclonal antibody

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Reactivity of an anti-(human gastric carcinoma) monoclonal antibody with core-related peptides of gastrointestinal mucin

Cited by 14 publications

References 16 publications

Monoclonal antibodies reacting with the MUC2 mucin core protein

Monoclonal antibodies reacting with the MUC2 mucin core protein

Mucin antibodies - new tools in diagnosis and therapy of cancer

Heteroclitic recognition of combinatorial TX1TX2T peptide mixtures by mucin‐2 protein specific monoclonal antibody

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Heteroclitic recognition of combinatorial TX¹TX²T peptide mixtures by mucin‐2 protein specific monoclonal antibody