Measurement of human granulocyte-macrophage colony-stimulating factor (GM-CSF) by enzyme-linked immunosorbent assay

Biotherapy. 1989;1(3):161-7. doi: 10.1007/BF02170885.

Abstract

An IgG monoclonal antibody against recombinant human granulocyte-macrophage colony-stimulating factor (GM-CSF), designated HGM1, was produced by fusion of immune mouse splenocytes with HAT-sensitive murine myeloma cells. A sandwich enzyme-linked immunosorbent assay (ELISA) for measurement of human GM-CSF was developed using this HGM1 and a polyclonal antibody against GM-CSF raised in a rabbit. GM-CSF in culture supernatants of phytohemagglutinin (PHA)- or concanavalin A (Con A)-stimulated peripheral blood mononuclear cells (PBMC) were measured by this ELISA system and the conventional CFU-GM colony formation method. The data indicated that the ELISA was highly efficient and sensitive for the detection of as little as 50 pg/ml recombinant GM-CSF. The CFU-GM colony assay may be influenced by other cytokines which can enhance or suppress colony formation, and ELISA for GM-CSF is more useful for kinetic studies of precise levels of production from PBMC.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal / biosynthesis
  • Colony-Forming Units Assay
  • Colony-Stimulating Factors / analysis*
  • Colony-Stimulating Factors / genetics
  • Colony-Stimulating Factors / immunology
  • Colony-Stimulating Factors / pharmacology
  • Enzyme-Linked Immunosorbent Assay
  • Female
  • Granulocyte Colony-Stimulating Factor
  • Granulocyte-Macrophage Colony-Stimulating Factor
  • Growth Substances / analysis*
  • Growth Substances / immunology
  • Humans
  • Leukocytes, Mononuclear / enzymology
  • Mice
  • Mice, Inbred BALB C
  • Neutrophils / metabolism
  • Recombinant Proteins / pharmacology

Substances

  • Antibodies, Monoclonal
  • Colony-Stimulating Factors
  • Growth Substances
  • Recombinant Proteins
  • Granulocyte Colony-Stimulating Factor
  • Granulocyte-Macrophage Colony-Stimulating Factor