- 作者: Albert M. Wu; June H. Wu; Hsiang-Wei Kuo; Anthony Herp;
- 作者服務機構: a Glyco-Immunochemistry Research Laboratory, Institute of Molecular and Cellular Biology, and; b Department of Microbiology and Immunology, Chang-Gung University, Kweishan, Taoyuan 333, Taiwan
- 中文摘要: --
- 英文摘要: The terminal a anomeric GalNAc residue is an essential sugar for the Tn glycotope, human blood group A determinant, and Forssman antigen. In a previous study [King M.J., Parson S.F., Wu A.M., Jones N., Transfusion 31: 142-149, 1991], we defined two monoclonal antibodies (MoAbs, BRIC66 and BRIC111) reacting with human Tn red blood cells. However, more advanced studies of these two MoAbs were hampered by the lack of availability of Gal/GalNAc related glycotopes. In order to use these antibodies as powerful probes to elucidate structural changes during life processes, we have characterized in detail the combining sites of these two MoAbs using enzyme-linked immunosorbent (ELISA) and inhibition assays with an extended glycan/ligand collection. From the results, it has been established that BRIC66 demonstrated multiple specificities and its reactivity towards glycotopes was defined as: GalNAcαl->Ser/Thr (Tn) > GalNAcαl->3(LFucαl->2)Gal (Ah) > GalNAcαl->3Galβ1->4Glc (AL) > GalNAαl->3Gal (A) > GalNAcαl-> 3GalNAc (F) > GalNAc >> Gal or Glc. Another MoAb, BRIO 11, mainly bound Tn-glycophorin. The best ligand for this MoAb was Tn-containing glycopeptides (M.W. < 3.0 x 10^3 Da) from asialo ovine salivary mucin (OSM), which was approximately 70 and 58 times more active than GalNAc and monomeric GalNAcαl->Ser/Thr (Tn), respectively, suggesting that the active glycotopes present in glycophorin for BRIC111 binding also exist in OSM. The N-acetyl group at carbon-2 and configuration at carbon-2 and carbon-4 of the α anomeric GalNAc are required for the binding of either MoAb. Identification of these binding properties should aid in the selection of these MoAbs and the conditions required for biological studies and clinical applications.
- 中文關鍵字: --
- 英文關鍵字: carbohydrate specificities, glycoprotein binding, monoclonal antibodies, T glycotopes, Tn epitopes