Preparation and Preliminary Application of the Monoclonal Antibodies to Recombinant Human Interferon-γ
Liu Yage; Pn Feifei;Cao Jianpihg; Shun Yihong (The general hospital of Shenyang Army,Shenyang 110011)
Four lines of hybridomas were prepared by the convenhonal method. The hybridomas,which were named 3D3, 3D12, 3F9 and 3B8 respectively, constantly secrete monoclonal antibodies(McAb)against recombinant human interferon-γ(IFN-γ).The subclasses of the McAbs were idenhfied by Ouchterlony test as IgGZb, IgG3,IgGI and IgGI respeCtly?ly.The relative affinity constantSfor the firSt two lines were 6.25×10-11 and those for latter two lines were 6.25x 10--13.The titers of theculture superatantS of the hybridomas measured by indirect ELISA were between Zx 102and l.28xlos. An of four McAbs purified from mouse ascites have cross-reactions to natural IFNry and effectSof neutralhation to recombinant human IFNry. The culture supernatantS of 3Fg and 3D3 also have aremarkable effeCt of neutralhation to the rocombinant human IFNry. The affinity chromatographiccolumn was prepared using 3F9 McAb and bromint cyan achvated Sepharose 4B, and was applied toPured crude IFNry. The purity of the IFNry after affinity chromatography is more than 95% and itSspeCific activity reach to 1.9×107IU / mg.