Identification of two potential receptor-binding sites of hGM-CSF

OGGERO EBERHARDT, M.; FRANK, R.; KRATJE, R. B.; ETCHEVERRIGARAY, M.

Transport Phenomena and Unit Operations ? Food Sience and Engineering Biotechnology

Centro de Publicaciones CERIDE-CONICET

Lugar: Santa Fe; Año: 2001 vol. 3 p. 1807 - 1812

1666-1621

Two receptor binding sites of hGM CSF are described. Competitive binding ELISA using four monoclonal antibodies (MAbs) showed their different epitope recognition. The antibody combining sites were mapped using sets of overlapping peptides and hexapeptide libraries prepared by the SPOT synthesis technique. We identified the conformational dependent epitopes A18E21R23R24F119 and R23E21N17W13 bound by MAb CC5B5 and the non linear epitope P118F119W13E14 bound by MAb M1B8. The epitopes recognized by these two MAbs are very closely located on the native protein surface. The peptide L61YKQGKLRGSLTK72 was recognized by MAb M7E10 and the peptide A1PAR4, representing the N terminal sequence of the protein, was bound by the non neutralizing MAb CC1H7. Inhibition assays of the GM CSF biological activity demonstrated that MAb M1B8, CC5B5 and M7E10 bind to domains which are responsible for the interaction of the cytokine with the GM CSF receptor. [Participants of the 3rd Mercosur Congress on Process Systems Engineering (ENPROMER) were invited to present a full paper, which was published in ?Transport Phenomena and Unit Operations ? Food Sience and Engineering Biotechnology?, ISSN 1666-1621, Centro de Publicaciones CERIDE-CONICET, Santa Fe (2001) Vol. III, p. 1807 -1812].