EGF cell surface receptor quantitation on ocular cells by an immunocytochemical flow cytometry technique

J. G. Lopez, Jin Chew Sek Jin Chew, H. W. Thompson, J. S. Malter, M. S. Insler, R. W. Beuerman

Research output: Contribution to journalArticle

34 Scopus citations

Abstract

A method is presented for the rapid flow cytometric determination of epidermal growth factor (EGF) receptor densities on the surface of cultured ocular cells. The technique uses a biotinylated monoclonal antibody directed against the EGF receptor in conjunction with a streptavidin-bound fluorochrome and requires the specific fluorescence per cell to be measured as a function of ligand and receptor concentration. Because the measurement is noninvasive and restricted to cell surface-bound material, the cells can be kept in a physiologic environment, even at the moment of assay. Calculated receptor densities ranged from 5142/cell (infant human corneal endothelium) to 35,678/cell (infant human keratocytes) to >5 x 105/cell for an A431 control cell line. Species and donor age differences were noted, as was transient receptor downregulation after EGF administration. Flow cytometry represents a valuable time saving procedure for large scale applications while providing the same level of sensitivity as standard radioimmunoassays. This technique is applicable to quantitation of other growth factor cell surface receptors and could greatly expand the use of flow cytometry in the research laboratory.

Original languageEnglish (US)
Pages (from-to)2053-2062
Number of pages10
JournalInvestigative Ophthalmology and Visual Science
Volume33
Issue number6
StatePublished - Jan 1 1992

Keywords

  • R-phycoerythrin
  • cell surface receptor
  • epidermal growth factor
  • flow cytometry
  • immunofluorescence
  • monoclonal antibody
  • streptavidin

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

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    Lopez, J. G., Sek Jin Chew, J. C., Thompson, H. W., Malter, J. S., Insler, M. S., & Beuerman, R. W. (1992). EGF cell surface receptor quantitation on ocular cells by an immunocytochemical flow cytometry technique. Investigative Ophthalmology and Visual Science, 33(6), 2053-2062.