title: Immunization with undenatured bovine zonular fibrils results in monoclonal antibodies to fibrillin.
abstract: Microfibrils were dissected from the zonular apparatus of the bovine eye, homogenized and used as an immunogen to prepare monoclonal antibodies. Initial screening of hybridomas was performed by immunoblotting to a sonicate of zonular fibrils and by immunolocalization to frozen sections of the zonular apparatus. Subsequently, monoclonal antibodies with strong immunoreactivity to zonular fibrils were shown to recognize microfibrils in a wide range of connective tissues both by immunofluorescent staining and by electron microscopic immunolocalization. All antibodies were found to recognize a single protein of 350 kDa on Western blotting of the proteins secreted by bovine aortic smooth muscle cells. A protein of the same molecular weight and properties was recognized by an antibody previously prepared by another group against fibrillin. A member of the fibrillin family therefore represents the major immunogen of intact zonular fibrils, and the results support previous evidence for a close relationship between zonular fibrils and other connective tissue microfibrils. The zonular apparatus is a suitable system to obtain purified preparations of microfibrils in order to investigate their composition and structural organization.
authors.affiliation: Department of Cell Biology, University of Alabama, Birmingham 35294.
authors.forename: D W
authors.initials: DW
authors.lastname: Wright
authors.forename: C N
authors.initials: CN
authors.lastname: McDaniels
authors.forename: S
authors.initials: S
authors.lastname: Swasdison
authors.forename: M A
authors.initials: MA
authors.lastname: Accavitti
authors.forename: P M
authors.initials: PM
authors.lastname: Mayne
authors.forename: R
authors.initials: R
authors.lastname: Mayne
mesh_terms: D008841:Actin Cytoskeleton / Q000276:immunology* / Q000648:ultrastructure; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D000917:Antibody Formation; D001011:Aorta / Q000166:cytology / Q000378:metabolism / Q000648:ultrastructure; D015153:Blotting, Western; D002417:Cattle; D002478:Cells, Cultured; D005123:Eye / Q000276:immunology*; D000071837:Fibrillins; D005455:Fluorescent Antibody Technique; D008840:Microfilament Proteins / Q000032:analysis* / Q000276:immunology*; D016253:Microscopy, Immunoelectron; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000378:metabolism / Q000648:ultrastructure
chemical_list: D000911:Antibodies, Monoclonal; D000071837:Fibrillins; D008840:Microfilament Proteins