Monoclonal antibody to fibulin-1 generated by genetic immunization

• Published in: Journal of cellular biochemistry 2003-07, Vol.89 (4), p.647-652
• Main Authors: Pupa, S.M., Forti, S., Invernizzi, A.M., Giovanazzi, R., Twal, W.O., Argraves, W.S., Ménard, Sylvie
• Format: Article
• Language: English
• Subjects: Adjuvants, Immunologic - genetics; Adjuvants, Immunologic - pharmacology; Animals; Antibodies, Monoclonal - biosynthesis; Antibodies, Monoclonal - genetics; Antibodies, Monoclonal - immunology; Antibodies, Monoclonal - isolation & purification; Breast Neoplasms - immunology; Breast Neoplasms - metabolism; Calcium-Binding Proteins - genetics; Calcium-Binding Proteins - immunology; Calcium-Binding Proteins - metabolism; Carcinoma, Squamous Cell - immunology; Carcinoma, Squamous Cell - ultrastructure; CpG oligodeoxyribonucleotides; Escherichia coli - genetics; Escherichia coli - metabolism; Extracellular Matrix Proteins - genetics; Extracellular Matrix Proteins - immunology; Extracellular Matrix Proteins - metabolism; Fibulin-1; genetic immunization; Humans; Hybridomas - immunology; Immunoglobulin Isotypes - chemistry; Immunoglobulin Isotypes - immunology; Immunoglobulin M - chemistry; Immunoglobulin M - immunology; Immunologic Tests - methods; Mice; monoclonal antibodies; Oligodeoxyribonucleotides - genetics; Oligodeoxyribonucleotides - immunology; Oligodeoxyribonucleotides - pharmacology; Plasmids - genetics; Recombinant Proteins - biosynthesis; Recombinant Proteins - genetics; Recombinant Proteins - immunology; Tumor Cells, Cultured
• ISSN: 0730-2312; 1097-4644
• DOI: 10.1002/jcb.10563

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‐1D 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. J. Cell. Biochem. 89: 647–652, 2003. © 2003 Wiley‐Liss, Inc.