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Monoclonal antibody to fibulin‐1 generated by genetic immunization
Author(s) -
Pupa S.M.,
Forti S.,
Invernizzi A.M.,
Giovanazzi R.,
Twal W.O.,
Argraves W.S.,
Ménard Sylvie
Publication year - 2003
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.10563
Subject(s) - monoclonal antibody , immunogen , microbiology and biotechnology , antibody , isotype , biology , immunoprecipitation , extracellular matrix , glycoprotein , breast carcinoma , breast cancer , cancer research , chemistry , cancer , immunology , genetics
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.