ELISA for quantitation of l-selectin shed from leukocytes in vivo

Olivier Spertini, Boris Schleiffenbaum, Cathy White-Owen, Phillip Ruiz, Thomas F. Tedder

Research output: Contribution to journalArticle

110 Scopus citations

Abstract

l-selectin is a cell surface receptor on granulocytes, lymphocytes and monocytes that is responsible for the initial attachment of leukocytes to endothelium. The extracellular domain of l-selectin is proteolytically shed from leukocytes following cellular activation in vitro. The shed form of l-selectin (sl-selectin) is functionally active and at high concentrations can inhibit leukocyte attachment to endothelium. Therefore, an ELISA was developed to quantitate the levels of sl-selectin in biological fluids, biopsy specimens and during recombinant protein production. This simple, quantitative sandwich ELISA uses two monoclonal antibodies directed against the extracellular domain of sl-selectin. The assay has a detection range of 5-1300 ng/ml, is precise and sensitive. The ability of this assay to detect sl-selectin in serum, plasma, and culture supernatant fluid was demonstrated and it was used to quantitate circulating sl-selectin in normal and patient sera. Patients with sepsis and HIV infection showed markedly elevated sl-selectin levels in serum. Thus, the ELISA should prove useful both for laboratory purposes as well as in the diagnostic evaluation of patients with inflammatory diseases.

Original languageEnglish (US)
Pages (from-to)115-123
Number of pages9
JournalJournal of Immunological Methods
Volume156
Issue number1
DOIs
StatePublished - Nov 25 1992

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Keywords

  • ELISA
  • l-Selectin
  • Leukocyte
  • Receptor
  • Shedding

ASJC Scopus subject areas

  • Biotechnology
  • Immunology

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