Process Optimization of PEGylating Fused Protein of LL-37 and Interferon-α2a
Author(s): ZHANG Mingjie, Pharmaceutical Co., Ltd.Guangdong Zijing Zhengtian
Pages: 1261-1266
Year: 2015 Issue: 6
Journal: Journal of Biomedical Engineering
Keyword: interferon-α2a; LL-37; polyethylene glycol(PEG); PEGylating;
Abstract: PEGylating is an effective way for prolonging the half-time period and decreasing the immunogenicity of protein drugs.With experiments of single factor,it was proved that the optimal processes for PEGylating the fused protein of LL-37 and interferon(IFN)-α2awere:PEG molecular weight was 5 000,fused protein concentration was0.6mg/mL,the mole ratio of protein to mPEG5000-SS was 1∶10,the reaction temperature was 4℃,and the pH was9.0,respectively.With orthogonal experiments,we proved that the influential order of 3main factors is:the fused protein concentration>the mole ratio of protein and mPEG5000-SS>pH and the optimal conditions were the fused protein concentration as 0.6mg/mL,the mole ratio of protein and mPEG5000-SS as 1∶10,pH as 8.8.Under these optimal conditions,the average rate of PEGylated protein with 3times parallel experiments was 86.98%.After PEGylated,the interferon activity and antimicrobial activity of fused protein could be remained higher than 58% and97%,respectively.
Pages: 1261-1266
Year: 2015 Issue: 6
Journal: Journal of Biomedical Engineering
Keyword: interferon-α2a; LL-37; polyethylene glycol(PEG); PEGylating;
Abstract: PEGylating is an effective way for prolonging the half-time period and decreasing the immunogenicity of protein drugs.With experiments of single factor,it was proved that the optimal processes for PEGylating the fused protein of LL-37 and interferon(IFN)-α2awere:PEG molecular weight was 5 000,fused protein concentration was0.6mg/mL,the mole ratio of protein to mPEG5000-SS was 1∶10,the reaction temperature was 4℃,and the pH was9.0,respectively.With orthogonal experiments,we proved that the influential order of 3main factors is:the fused protein concentration>the mole ratio of protein and mPEG5000-SS>pH and the optimal conditions were the fused protein concentration as 0.6mg/mL,the mole ratio of protein and mPEG5000-SS as 1∶10,pH as 8.8.Under these optimal conditions,the average rate of PEGylated protein with 3times parallel experiments was 86.98%.After PEGylated,the interferon activity and antimicrobial activity of fused protein could be remained higher than 58% and97%,respectively.
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