Monoclonal antibody to fibulin-1 generated by genetic immunization.
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.
Pupa
S M
SM
Molecular Targeting Unit, Department of Experimental Oncology, Istituto Nazionale Tumori, Milan, Italy.
Forti
S
S
Invernizzi
A M
AM
Giovanazzi
R
R
Twal
W O
WO
Argraves
W S
WS
Ménard
Sylvie
S
D000276:Adjuvants, Immunologic / Q000235:genetics / Q000494:pharmacology; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology* / Q000302:isolation & purification; D001943:Breast Neoplasms / Q000276:immunology / Q000378:metabolism; D002135:Calcium-Binding Proteins / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism; D002294:Carcinoma, Squamous Cell / Q000276:immunology / Q000648:ultrastructure; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D016326:Extracellular Matrix Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D007132:Immunoglobulin Isotypes / Q000737:chemistry / Q000276:immunology; D007075:Immunoglobulin M / Q000737:chemistry / Q000276:immunology; D007159:Immunologic Tests / Q000379:methods; D051379:Mice; D009838:Oligodeoxyribonucleotides / Q000235:genetics / Q000276:immunology / Q000494:pharmacology; D010957:Plasmids / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D014407:Tumor Cells, Cultured
D000276:Adjuvants, Immunologic; D000911:Antibodies, Monoclonal; D002135:Calcium-Binding Proteins; D016326:Extracellular Matrix Proteins; D007132:Immunoglobulin Isotypes; D007075:Immunoglobulin M; D009838:Oligodeoxyribonucleotides; D011994:Recombinant Proteins; C060820:fibulin