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-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.
Copyright 2003 Wiley-Liss, Inc.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Adjuvants, Immunologic / genetics
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Adjuvants, Immunologic / pharmacology
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Animals
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Antibodies, Monoclonal / biosynthesis
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Antibodies, Monoclonal / genetics*
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Antibodies, Monoclonal / immunology*
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Antibodies, Monoclonal / isolation & purification
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Breast Neoplasms / immunology
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Breast Neoplasms / metabolism
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Calcium-Binding Proteins / genetics*
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Calcium-Binding Proteins / immunology*
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Calcium-Binding Proteins / metabolism
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Carcinoma, Squamous Cell / immunology
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Carcinoma, Squamous Cell / ultrastructure
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Escherichia coli / genetics
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Escherichia coli / metabolism
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Extracellular Matrix Proteins / genetics
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Extracellular Matrix Proteins / immunology
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Extracellular Matrix Proteins / metabolism
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Humans
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Hybridomas / immunology
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Immunoglobulin Isotypes / chemistry
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Immunoglobulin Isotypes / immunology
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Immunoglobulin M / chemistry
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Immunoglobulin M / immunology
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Immunologic Tests / methods
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Mice
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Oligodeoxyribonucleotides / genetics
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Oligodeoxyribonucleotides / immunology
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Oligodeoxyribonucleotides / pharmacology
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Plasmids / genetics
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Recombinant Proteins / biosynthesis
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Recombinant Proteins / genetics
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Recombinant Proteins / immunology
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Tumor Cells, Cultured
Substances
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Adjuvants, Immunologic
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Antibodies, Monoclonal
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Calcium-Binding Proteins
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Extracellular Matrix Proteins
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Immunoglobulin Isotypes
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Immunoglobulin M
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Oligodeoxyribonucleotides
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Recombinant Proteins
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fibulin