摘要
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。
Comparisons between native and PEGylated consensus interferon α on satiabbility were maae as well as the measurement and analysis of molecular weight were conducted. The thermal stability, the enzymolysis stability and the stability in human serum in vitro of PEG-IFNcon and native IFNcon were evaluated. It also tested the anti-viral activity of IFNeon, monoPEG-IFNcon and oligoPEG-IFNcon. Relative Mr of PEG-IFNcon and native IFNcon was detected by MALDI TOF-MS and then was analysised by SE HPLC and SDS-PAGE. The thermal stability, stability in serum and trypsin solution of PEG-IFNcon were significantly more stable than those of the IFNcon. The anti-viral activity of IFNcon, monoPEG-IFNcon and oligoPEG-IFNcon were 6.72× 10^8 8. 1 × 10^7 , 4. 0× 10^6 IU/mg respectively. Relative Mr of PEC-IFNcon was 62665 and native IFNcon was 19613 detected by MALDI-TOF- MS. The apparent Mr measured by SD-PAGE was about 100 k. Retention time of PEG-IFNcon detected by SE HPLC was 21. 260 rain and IFNcon was 28. 855 rain. Compared with IFNcon, PEG-IFNcon exhibits on improved stability and this may be helpful clinically for the research of medicine.
出处
《药物生物技术》
CAS
CSCD
2009年第1期28-32,共5页
Pharmaceutical Biotechnology
基金
国家高技术研究发展计划(863计划)项目(2007AA02Z101)
国家自然科学基金(30672559)
教育部"新世纪优秀人才支持计划"项目(NCET-04-0506)
国家基础科学人才培养基金(J0630858)