Generation and characterization of anti-MUC4 monoclonal antibodies reactive with normal and cancer cells in humans.

Moniaux, N and Varshney, Grish C and Chauhan, Subhash Chand and Copin, Marie Christine and Jain, M and Wittel, Uwe A and Andrianifahanana, Mahefatiana and Aubert, Jean-Pierre and Batra, Surinder K (2004) Generation and characterization of anti-MUC4 monoclonal antibodies reactive with normal and cancer cells in humans. The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society, 52 (2). pp. 253-61. ISSN 0022-1554

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We have previously cloned the full-length cDNA (approximately 28 Kb) and established the complete genomic organization (25 exons/introns over 100 kb) of the human MUC4 mucin. This large molecule is predicted to protrude over 2 microm above the cell surface, in which MUC4alpha is an extracellular mucin-type glycoprotein subunit and MUC4beta is the transmembrane subunit. Over two thirds of the encoded protein sequence consists of 16-amino-acid tandem repeats (TR), which are flanked by unique sequences. In this study we generated and characterized monoclonal antibodies (MAbs) directed against the TR region of MUC4. Mice were immunized with a KLH-conjugated MUC4 TR peptide, STGDTTPLPVTDTSSV. Several clones were purified by three rounds of limited dilutions and stable clones presenting a sustained antibody production were selected for subsequent characterization. Antibodies were tested for their reactivity and specificity to recognize the MUC4 peptide and further screened by enzyme-linked immunosorbent assay (ELISA) and Western blotting analyses. One of the MAbs (8G7) was strongly reactive against the MUC4 peptide and with native MUC4 from human tissues or pancreatic cancer cells in Western blotting, immunohistochemistry, and confocal analysis. Anti-MUC4 MAb may represent a powerful tool for the study of MUC4 function under normal and pathological conditions and for diagnosis of solid tumors including those in the breast, pancreas, lungs, and ovaries.

Item Type: Article
Additional Information: Copyright of this article belongs to Sage Publication/ The Histochemical Society.
Uncontrolled Keywords: mucin; MUC4; antibody; pancreatic adenocarcinoma
Subjects: Q Science > QD Chemistry
Depositing User: Dr. K.P.S.Sengar
Date Deposited: 14 Feb 2012 10:31
Last Modified: 09 Jan 2015 08:50

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