Identification of functionally distinct domains of human granulocyte- macrophage colony-stimulating factor using monoclonal antibodies

摘要: Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a glycoprotein that required for the survival, growth, and differentiation of hematopoietic progenitor cells. Although primary structure GM-CSF known from cDNA cloning, relationship between function not fully understood. Fifteen different monoclonal antibodies (MoAbs) to human were generated map immunologically distinct areas molecule. Each MoAbs was biotinylated shown by enzyme-linked immunosorbent assay bind recombinant had been affixed solid phase. 15 unconjugated then used compete with each MoAb binding GM-CSF. These cross-blocking studies identified eight epitopes native Seven these also present in denatured Western blotting, four at least partially conserved on reduced beta-mercaptoethanol. neutralized both (glycosylated nonglycosylated) natural GM colony-forming unit (CFU-GM) blocked GM-CSF-induced activation neutrophils. For most there good correlation neutralizing activity capacity block 125I-GM-CSF neutrophils or blasts. Non-neutralizing one epitope None interleukin-3, G-CSF, M-CSF. The locations seven could be mapped regard amino acid determining reactivity synthetic peptides human-mouse chimeric GM-CSFs. found acids 40–77, 78–94, 110–127. Thus, are useful identify functional domains identifying regions likely involved receptor interaction.