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Interferon-gamma(IFN-γ)was induced by pokeweed mitogen(PWM)or esculentoside in human splenic lymphocytes.PWM-inducedIFN-γ was partially purified to a specific activity of 10~6 RU/mgprotein by batch adsorption on controlled-pore glass(CPG)beadsand desorption by ethylene glycol in phosphate-buffered saline(PBS).Use of Chinese made micro-pore glass(MPG)instead of CPG topurify IFN-γ showed similar purification efficacy.Both CPG andMPG recovered could be regenerated for repeated use.The yieldfrom glass beads was further purified to a specific activity of 10~7RU/mg protein,with about 78% recovery,by the anti-new-borncalf serum antibodies-Sepharose 4B(ACS)column chromatography.Almost all the antiviral activity of the IFN-γ induced by PWM oresculentoside was destroyed by adjusting to pH 2 or heating to 56℃,and was neutralized by monoclonal antibody to human interferon-γ(Hu IFN-γ),thereby the product was identified as Hu IFN-γ.Bio-gel P-200 chromatography resulted in two peaks of activity for PWM-induced IFN,a major one with molecular weight(MW)of 40,000-45,700 and a minor one of 25,700,but only one peak of 45,700-49,000for esculentoside-induced IFN.
Interferon-gamma was induced by pokeweed mitogen (PWM) or esculentoside in human splenic lymphocytes. PWM-inducedIFN-γ was partially purified to a specific activity of 10 ~ 6 RU / mg protein by batch adsorption on controlled-pore glass (CPG) beadsand desorption by ethylene glycol in phosphate-buffered saline (PBS). Use of Chinese made micro-pore glass (MPG) instead of CPG topurify IFN-γ showed similar purification efficacy. The yield from glass beads was further purified to a specific activity of 10-7 RU / mg protein with about 78% recovery by the anti-new-borncalf serum antibodies-Sepharose 4B (ACS) column chromatography. Almost all the antiviral activity of the IFN-γ induced by PWM oresculentoside was destroyed by adjusting to pH 2 or heating to 56 ° C and was neutralized by monoclonal antibody to human interferon-γ (Hu IFN-γ), thus the product was identified as Hu IFN-γ. Bio-gel P-200 chromatography resulted in two peaks of activ ity for PWM-induced IFN, a major one with molecular weight (MW) of 40,000-45,700 and a minor one of 25,700, but only one peak of 45,700-49,000 for esculentoside-induced IFN.