Mapping of binding sites for monoclonal antibodies to chick tropoelastin by recombinant DNA techniques.
A fusion molecule consisting of the entire coding sequence of mature chicken tropoelastin preceded by 14 amino acids of the signal peptide and 9 amino acids of vector origin has been expressed in a recombinant bacterial system and purified. The molecule has been used as immunogen for the production of hybridomas. Monoclonal antibodies which bound specifically the immunogen were also reactive with tropoelastin purified from chick aorta and stained elastic fibers in aorta sections by immunofluorescence. The region of tropoelastin containing the antigenic determinant recognized by each antibody has been identified by a recombinant DNA expression strategy based on the use of cDNA clones spanning different portions of the coding sequence. It could be shown that several antibodies were directed against unique epitopes; among these, a group of antibodies bound specifically to the sequence (PGVGV)n. Other antibodies were found to recognize antigenic determinants present more than once in the molecule. The monoclonal antibodies thus characterized will be useful reagents in studying the function of the different domains of tropoelastin.
Marigo
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V
Institute of Histology and Embryology, University of Padova, Italy.
Sitta
A
A
Volpin
D
D
Bressan
G M
GM
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis* / Q000276:immunology; D000918:Antibody Specificity; D001666:Binding Sites, Antibody; D002642:Chick Embryo; D002874:Chromosome Mapping; D004247:DNA; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes; D006825:Hybridomas / Q000276:immunology; D051379:Mice; D011994:Recombinant Proteins; D013045:Species Specificity; D014333:Tropoelastin / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000939:Epitopes; D011994:Recombinant Proteins; D014333:Tropoelastin; D004247:DNA