A new alternative for simultaneous immunohistochemical screening of 96 hybridoma clones for tissue-specific antibody productions selects a monoclonal antibody to insect corpus cardiacum.
In the current search for the elucidation of the true structure of hitherto unidentified 'new' insect neuropeptides we designed a novel screening method to facilitate the primary detection of neurone-specific antibody secreting mouse-mouse hybridoma clones obtained after immunization with neuronal tissue homogenates. The present procedure is principally adapted from a conventional immunohistological test and enables one to rapidly screen 96 (and even more) clones at one time for potential secretion of specific antibodies to different tissue compounds, without the necessity of having a purified antigen. It has proved to be sensitive, rapid, practical and reproducible. As such it promises to be very useful to discriminate amongst the wide range of antibodies to various kinds of materials produced by hybridomas by detecting monoclonal antibodies directed against factors contained in well-defined tissues in which one is interested. This paper also reports the successful application of this method to a primary screening of clones producing murine monoclonal antibodies to substances of insect corpora cardiaca (CC), after immunization with crude antigen preparations.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000276:immunology*; D000917:Antibody Formation; D000918:Antibody Specificity; D002999:Clone Cells; D003058:Cockroaches; D006110:Grasshoppers; D006651:Histocytochemistry / Q000379:methods*; D006825:Hybridomas / Q000276:immunology*; D007120:Immunochemistry / Q000379:methods*; D007313:Insecta / Q000276:immunology*; D009490:Neurosecretory Systems / Q000276:immunology*
D000911:Antibodies, Monoclonal