PARTIAL PURIFICATION AND PHYSICOCHEMICAL CHARACTERIZATION OF HUMAN SNTERFERON-?
- VernacularTitle:人?干扰索的纯化及理化性质的研究
- Author:
Gensheng FENG
- Publication Type:Journal Article
- Keywords:
interferon purification pokeweed mitogen esculentoside
- From:
Academic Journal of Second Military Medical University
1982;0(02):-
- CountryChina
- Language:Chinese
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Abstract:
Interferon-gamma(IFN-?)was induced by PWM or esculentoside in human splenic lymphocytes. PWM-induced IFN-r was partially purified to a spe. act. of 106 RU/mg protein by batch adsorption on controlled-pore glass (CPG) beads and desorption by ethylene glycol in PBS.Use of native-made micro-pore glass(MPG)instead of CPG to purify IFN-r showed similar purification efficacy.Both CPG and MPG recovered could be regenerated for repeated use.The yield from glass beads was further purified to a spe.act.of 107 RU/mg protein, with about 78%recovery, by the anti-new-born calf serum antibodies-Sepharose 4B(ACS)column chromatography.Almost all the antiviral activity induced by PWM or esculentoside was destroyed by pH 2 or heat at 56℃ and neutralized by monoclonal antibody against HuIFN-r, thereby identified as HuIFN-r. Bio-Gel P-200 chromatography resulted in two peaks of activity for PWM-induced IFN, a major one with molecular weight of 40,000-45,700 and a minor one of 25,700 dalton, but only one peak with molecular weight of 45,700-49,000 for esculentoside-induced IFN.
