Production and characterization of murine monoclonal antibodies to polypeptide hormones and their fragments.
The diagnostic importance of monoclonal antibodies assays for peptide hormones in biological fluids and for histological peptide localization is rapidly increasing. In our laboratory a general protocol has been developed for immunization and fusion that has been proven very useful, with minor modifications, for generating monoclonal antibodies against insulin, calcitonin, adrenocorticotropin and parathyroid hormone. Our procedure offers the following advantages: 1) it requires a relatively low amount of antigen; 2) it takes only 16-20 days from the first immunization to the time of fusion; 3) it mostly generates IgG producing hybrids; 4) it involves few manipulations of the animals and no i.v. injections. The widely used methods utilized for peptide carrier conjugation, few guidelines for the choice of peptide fragments suitable as immunogens and some applications of antipeptide monoclonal antibodies will be briefly discussed.
Modena
D
D
Racchetti
G
G
Putignano
S
S
Comitti
R
R
Galante
Y M
YM
D000324:Adrenocorticotropic Hormone / Q000276:immunology; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology* / Q000302:isolation & purification; D002116:Calcitonin / Q000276:immunology; D002459:Cell Fusion; D006241:Haptens / Q000276:immunology; D006728:Hormones / Q000276:immunology*; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D007114:Immunization; D007118:Immunoassay; D007150:Immunohistochemistry; D051379:Mice; D010281:Parathyroid Hormone / Q000276:immunology; D010446:Peptide Fragments / Q000276:immunology; D010455:Peptides / Q000276:immunology*
D000911:Antibodies, Monoclonal; D006241:Haptens; D006728:Hormones; D010281:Parathyroid Hormone; D010446:Peptide Fragments; D010455:Peptides; D000324:Adrenocorticotropic Hormone; D002116:Calcitonin