Monoclonal antibody to fibulin-1 generated by genetic immunization

S. M. Pupa, S. Forti, A. M. Invernizzi, R. Giovanazzi, W. O. Twal, W. S. Argraves, Sylvie Ménard

Research output: Contribution to journalArticlepeer-review

Abstract

Fibulin-1 (Fbln-1) is an extracellular matrix (ECM) and plasma glycoprotein. Considering the growing evidence indicating that Fbln-1 plays a role in cancer we sought to develop monospecific antibodies to better facilitate further studies of the function of Fbln-1 in breast cancer. Using a plasmid expression vector encoding full-length human Fbln-1 D as an immunogen and CpG oligodeoxyribonucleotides as adjuvant a monoclonal antibody (MAb) against Fbln-1 was produced. This MAb, designated MEM-2 was of IgM isotype and reacted with bacterially expressed Fbln-1. Furthermore, MEM-2 reacted with Fbln-1 expressed in the ECM released by cultured human breast carcinoma SKBR-3 cells in ELISA, and also with Fbln-1 present in SKBR-3 cell extract in immunoprecipitation and Western blotting. MEM-2 also reacted with Fbln-1 in human breast carcinoma specimens. These findings illustrate the utility of genetic immunization as a means of generating monoclonal antibodies to tumor-related ECM proteins. MEM-2 represents a useful new tool for the study of Fbln-1 in breast cancer.

Original languageEnglish
Pages (from-to)647-652
Number of pages6
JournalJournal of Cellular Biochemistry
Volume89
Issue number4
DOIs
Publication statusPublished - Jul 1 2003

Keywords

  • CpG oligodeoxyribonucleotides
  • Fibulin-1
  • Genetic immunization
  • Monoclonal antibodies

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology

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