Human granulocyte-macrophage colony-stimulating factor (hGM-CSF): Identification of a binding site for a neutralizing antibody

Edouard Nice, Peter Dempsey, Judith Layton, George Morstyn, Da Fu Cup, Richard Simpson, Louis Fabri, Antony Burgess

Research output: Contribution to journalArticleResearchpeer-review

14 Citations (Scopus)


One approach to the localization of functionally active regions of human granulocyte-macrophage colony-stimulating factor (hGM-CSF) is to map the epitopes recognized by neutralizing anti-hGM-CSF monoclonal antibodies. We have defined the epitope recognized by one neutralizing antibody (LMM102) using proteolytic fragments obtained by enzymic digestion of bacterially synthesized hGM-CSF. RP-HPLC fractionation of a tryptic digest resulted in the identification of an immunoreactive "tryptic core" peptide containing 66 amino acids (52% of the protein). Further digestion of this "tryptic core" with S. aureus V8 protease produced a unique immunoreactive hGM-CSF product comprising two peptides, residues 86-93 and 112-127, linked by a disulfide bond between residues 88 and 121. The individual peptides, generated by reduction with dithiothreitol, were not recognized by the antibody. An analog of this peptide has been synthesized chemically and shown to have similar immunoreactivity to the epitope obtained by enzymic digestion. A series of modified peptides has also been synthesized to identify further the region required for antibody recognition.

Original languageEnglish
Pages (from-to)159-169
Number of pages11
JournalGrowth Factors
Issue number2
Publication statusPublished - 1 Jan 1990
Externally publishedYes


  • Epitope mapping
  • GM-CSF

Cite this