Products of cells from gliomas: VIII. Multiple-well immunoperoxidase assay of immunoreactivity of primary hybridoma supernatants with human glioma and brain tissue and cultured glioma cells

P. E. McKeever, R. L. Wahl, P. Shakui, G. A. Jackson, L. H. Letica, M. Liebert, J. A. Taren, W. H. Beierwaltes, J. T. Hoff

Research output: Contribution to journalArticlepeer-review

Abstract

To test the feasibility of primary screening of hybridoma supernatants against human glioma tissue, over 5000 combinations of hybridoma supernatants with glioma tissue, cultured glioma cells, and normal central neural tissue were screened with a new multiple-well (M-well) screening system. This is an immunoperoxidase assay system with visual endpoints for screening 20-30 hybridoma supernatants per single microsope slide. There were extensive differences between specificities to tissue and to cultured glioma cells when both were screened with M-wells and when cultured cells were screened with standard semi-automated fluorescence. Primary M-well screening with glioma tissue detected seven hybridoma supernatants that specifically identified parenchymal cells of glioma tissue and that were not detected with cultured cells. Immunoreactivities of individual supernatants for vascular components (nine supernatants), necrosis (five supernatants), and nuclei (three supernatants) were detected. Other supernatants bound multiple sites on glioma tissue and/or subpopulations of neurons and glia of normal tissue. The results show that primary screening with glioma tissue detects a number of different specificities of hybridoma supernatants to gliomas not detected by conventional screening with cultured cells. These are potentially applicable to diagnosis and therapy.

Original languageEnglish
Pages (from-to)815-822
Number of pages8
JournalJournal of Histochemistry and Cytochemistry
Volume38
Issue number6
DOIs
StatePublished - 1990

Keywords

  • brain tissue
  • cell compartments
  • human glioma
  • hybridoma screening
  • immunoperoxidase
  • multiple-well screening
  • primary specificity

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