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A novel monoclonal antibody for detection of folate receptor alpha in paraffin-embedded tissues

Lookup NU author(s): Professor Alan Calvert, Professor John LunecORCiD

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Abstract

Folate biochemical pathway components such as FR-α are determinants of response to novel antifolate drugs such as pemetrexed. Knowledge of their level of expression in tumors will enable their optimal use by identifying potentially responsive subgroups of patients. In spite of its importance in the diagnosis and treatment of cancer, monoclonal antibodies to FR-α suitable for immunohistochemical analysis of formalin-fixed and paraffin-embedded biopsy samples, or that can be used for Western blot analysis, are not available. The aim of this study was thus to generate a monoclonal antibody that could be used to detect specific expression of FR-α in paraffin-embedded tissues. A 189 amino acid region of the FR-α sequence was expressed as a recombinant fusion protein and used as antigen to generate monoclonal antibodies. Studies by ELISA and Western blot identified specific reactivity of the BN3.2 antibody to the recombinant protein and a single 40kD protein in whole cell lysates from cell lines known to express FR-α. Immunohistochemical analysis of FR-α using hybridoma clone BN3.2 in a panel of normal tissues demonstrated expression limited to ovarian epithelia, placental trophoblasts, and proximal kidney tubules. Analysis of a panel of malignant and benign tissues demonstrated limited expression with variable intensities of staining and patterns of both membrane and cytoplasmic reactivity observed between cases. In the majority of malignant ovarian tumors, high intensity staining was observed, predominantly localized to the plasma membrane. Our results show that clone BN3.2 is a sensitive tool for detection of FR-α in paraffin-embedded tissues. This preliminary study also supports its use in immunohistochemical studies to determine the role of FR-α as a tumor prognostic marker and a possible therapeutic target. © 2007 Mary Ann Liebert, Inc.

Publication metadata

Author(s): Smith, A., Pinkney, M., Piggott, N., Calvert, A. H., Milton, I., Lunec, J.

Publication type: Article

Publication status: Published

Journal: Hybridoma

Year: 2007

Volume: 26

Issue: 5

Pages: 281-288

Print publication date: 01/10/2007

ISSN (print): 1554-0014

ISSN (electronic): 1557-8348

URL: http://dx.doi.org/10.1089/hyb.2007.0512

DOI: 10.1089/hyb.2007.0512

PubMed id: 17979543

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