Differential binding characteristics of protein G and protein A for Fc fragments of papain-digested mouse IgG


Aybay C.

IMMUNOLOGY LETTERS, cilt.85, sa.3, ss.231-235, 2003 (SCI-Expanded) identifier identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 85 Sayı: 3
  • Basım Tarihi: 2003
  • Doi Numarası: 10.1016/s0165-2478(02)00262-6
  • Dergi Adı: IMMUNOLOGY LETTERS
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Sayfa Sayıları: ss.231-235
  • Anahtar Kelimeler: mouse IgG, papain digestion, Fab fragment, Fc fragment, staphylococcal protein A, streptococcal protein G, affinity purification, MONOCLONAL-ANTIBODIES, AFFINITY-CHROMATOGRAPHY, STAPHYLOCOCCUS-AUREUS, RHEUMATOID FACTORS, CRYSTAL-STRUCTURE, IMMUNOGLOBULIN-G, FAB FRAGMENT, REGION, RECEPTORS, SUPERNATANTS
  • Gazi Üniversitesi Adresli: Hayır

Özet

It has been previously reported that staphylococcal protein A (SPA) bound only to the Fc region of mouse immunoglobulin G. (IgG) and streptococcal protein G (SPG) bound to both Fab and Fc regions of mouse IgG and the binding sites for SPG and SPA on Fc were overlapped. In this study the binding characteristics of SPG and SPA for papain-digested mouse IgG were analysed. Papain digestion of mouse IgG purified from CAy-IFNg99C hybridoma (secreting IgG1 monoclonal antibody specific for human interferon gamma)-induced ascites resulted in Fab and two major Fe fragments referred to as the high molecular weight (HMW) and the low molecular weight (LMW) Fc fragments. SPG bound to Fab and the LMW Fc fragments of the papain-digested IgG. However SPG did not bind to the HMW Fc fragment. SPA showed practically no reactivity with the Fab and the LMW Fc fragments of the papain-digested mouse IgG but only to the HMW Fc fragment. SPG and SPA binding assays showed that papain digestion discriminated the SPG and SPA binding sites in the Fc fragment of mouse IgG. These results demonstrated a clear evidence for the presence of two independent SPG and SPA binding sites in the Fc fragment of mouse IgG. (C) 2002 Elsevier Science B.V. All rights reserved.