title: Production of monoclonal antibodies to thyroglobulin by in vitro immunization with a free synthetic peptide.
abstract: A synthetic peptide corresponding to amino acids 1-19 of thyroglobulin was used to test the possibility of generating protein-reactive monoclonal antibodies by immunization in vitro with a synthetic peptide as antigen. Splenocytes from non-immunized Balb/c mice were cultured in serum-free medium for 3 days in the presence of thymocyte-conditioned medium and the synthetic peptide prior to fusion with SP2/0 murine myeloma cells. The synthetic peptide was used in its free form, i.e. not coupled to a protein carrier. Hybridomas secreting monoclonal antibodies reactive with the synthetic peptide were obtained after immunization in vitro with as little as 10 ng/ml of the synthetic peptide. Between 50 and 70% of the primary clones obtained in different experiments produced monoclonal antibodies also reactive with the intact protein. Six stable hybridomas were isolated; all produced antibodies of the IgM class. We conclude that immunization in vitro with a free synthetic peptide is an efficient method for the generation of monoclonal antibodies reactive with the intact protein.
authors: Laboratory of Biochemistry, Biotechnology Centre, University of Amsterdam, The Netherlands.
authors: M
authors: M
authors: de Boer
authors: F A
authors: FA
authors: Ossendorp
authors: B J
authors: BJ
authors: Al
authors: J
authors: J
authors: Hilgers
authors: J J
authors: JJ
authors: de Vijlder
authors: J M
authors: JM
authors: Tager
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D000918:Antibody Specificity; D001324:Autoantigens / Q000276:immunology; D006825:Hybridomas / Q000276:immunology; D007075:Immunoglobulin M / Q000096:biosynthesis; D051379:Mice; D008807:Mice, Inbred BALB C; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000276:immunology*; D013154:Spleen / Q000276:immunology; D013954:Thyroglobulin / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D001324:Autoantigens; D007075:Immunoglobulin M; D010446:Peptide Fragments; D013954:Thyroglobulin