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比较重组复合α干扰素(IFNcon)与经PEG修饰后产物的初步稳定性,测定其分子质量并进行分析。对IFNcon与PEG修饰IFNcon(PEG-IFNcon)进行60℃热稳定性,抗胰蛋白酶能力和人血清中稳定性的考察;比较IFNcon、monoPEG-IFNcon和oligoPEG-IFNcon的抗病毒生物活性;采用基质辅助激光解吸附飞行时间质谱(MALDI-TOF-MS)测定IFNcon和PEG-IFNcon的Mr;并利用SE-HPLC和SDS-PAGE对修饰产物进行分析。PEG-IFNcon的稳定性高于IFNcon;IFNcon、monoPEG-IFNcon和oligoPEG-IFNcon的抗病毒生物活性分别为6.72×108,8.1×107,4.0×106IU/mg;MALDI-TOF-MS测定IFNcon及PEG-IFNcon的Mr分别19613及62665;SDS-PAGE测得PEG-IFNcon表观Mr约100k;SE-HPLC测定IFNcon及PEG-IFNcon保留时间为28.855min和21.260min。经修饰,IFNcon稳定性显著提高,为该药物的临床研究提供理论基础。
The preliminary stability of the recombinant interferon alpha (IFNcon) and PEG-modified products was compared and the molecular mass was determined and analyzed. The thermal stability, antitrypsin ability and stability in human serum at 60 ° C of IFNcon and PEG-modified IFNcon (PEG-IFNcon) were examined; the antiviral biological activity of IFNcon, monoPEG-IFNcon and oligoPEG-IFNcon was compared; The Mrcons of IFNcon and PEG-IFNcon were determined by laser desorption / emission time of flight mass spectrometry (MALDI-TOF-MS). The modified products were analyzed by SE-HPLC and SDS-PAGE. The antiviral biological activity of IFNcon, monoPEG-IFNcon and oligoPEG-IFNcon were 6.72 × 108, 8.1 × 107 and 4.0 × 106IU / mg, respectively. The MALDI-TOF- IFNcon Mr respectively 19613 and 62665; SDS-PAGE measured apparent PEG-IFNcon Mr about 100k; SE-HPLC determination of IFNcon and PEG-IFNcon retention time of 28.855min and 21.260min. Modified, IFNcon stability was significantly improved, providing a theoretical basis for the clinical research of the drug.