Generation and Characterization of Anti-MUC4 Monoclonal Antibodies Reactive with Normal and Cancer Cells in Humans

Nicolas Moniaux, Grish Chandra Varshney, Subhash Chauhan, Marie Christine Copin, Maneesh Jain, Uwe A. Wittel, Mahefatiana Andrianifahanana, Jean Pierre Aubert, Surinder Kumar Batra

Research output: Contribution to journalArticle

66 Citations (Scopus)

Abstract

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 μm above the cell surface, in which MUC4α is an extracellular mucin-type glycoprotein subunit and MUC4β 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, STGDT-TPLPVTDTSSV. 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.

Original languageEnglish (US)
Pages (from-to)253-261
Number of pages9
JournalJournal of Histochemistry and Cytochemistry
Volume52
Issue number2
DOIs
StatePublished - Jan 1 2004
Externally publishedYes

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Tandem Repeat Sequences
Monoclonal Antibodies
Mucins
Peptides
Clone Cells
Western Blotting
Neoplasms
Pancreatic Neoplasms
Introns
Antibody Formation
Pancreas
Ovary
Exons
Glycoproteins
Breast
Complementary DNA
Enzyme-Linked Immunosorbent Assay
Immunohistochemistry
Amino Acids
Lung

All Science Journal Classification (ASJC) codes

  • Anatomy
  • Histology

Cite this

Generation and Characterization of Anti-MUC4 Monoclonal Antibodies Reactive with Normal and Cancer Cells in Humans. / Moniaux, Nicolas; Varshney, Grish Chandra; Chauhan, Subhash; Copin, Marie Christine; Jain, Maneesh; Wittel, Uwe A.; Andrianifahanana, Mahefatiana; Aubert, Jean Pierre; Batra, Surinder Kumar.

In: Journal of Histochemistry and Cytochemistry, Vol. 52, No. 2, 01.01.2004, p. 253-261.

Research output: Contribution to journalArticle

Moniaux, N, Varshney, GC, Chauhan, S, Copin, MC, Jain, M, Wittel, UA, Andrianifahanana, M, Aubert, JP & Batra, SK 2004, 'Generation and Characterization of Anti-MUC4 Monoclonal Antibodies Reactive with Normal and Cancer Cells in Humans', Journal of Histochemistry and Cytochemistry, vol. 52, no. 2, pp. 253-261. https://doi.org/10.1177/002215540405200213
Moniaux, Nicolas ; Varshney, Grish Chandra ; Chauhan, Subhash ; Copin, Marie Christine ; Jain, Maneesh ; Wittel, Uwe A. ; Andrianifahanana, Mahefatiana ; Aubert, Jean Pierre ; Batra, Surinder Kumar. / Generation and Characterization of Anti-MUC4 Monoclonal Antibodies Reactive with Normal and Cancer Cells in Humans. In: Journal of Histochemistry and Cytochemistry. 2004 ; Vol. 52, No. 2. pp. 253-261.
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