目的 对重组人生长激素-Fc(recombinant human growth hormone-Fc,rhGH-Fc)融合蛋白中试工艺的病毒灭活/去除效果进行验证,以期保证制品质量及患者安全.方法 选择伪狂犬病病毒(pseudorabies virus,PRV)、异嗜性小鼠白血病病毒(xenotropic murine leukemia virus,X-MuLV)、小鼠微小病毒(minute virus of mice,MVM)、呼肠孤病毒 Ⅲ 型(reovirus-3,Reo-3)4种病毒作为指示病毒,在缩小规模的条件下考察有机溶剂/表面活性剂(solvent and detergent,S/D)法对PRV、X-MuLV的灭活能力;亲和层析法对X-MuLV、MVM的去除能力;膜过滤法对MVM、Reo-3的去除能力.结果 S/D法对PRV和X-MuLV的灭活能力分别≥4.95、＞5.15 Log;亲和层析法对X-MuLV和MVM的去除能力分别≥4.36、1.68 Log;膜过滤法对MVM和Reo-3的去除能力分别＞5.36、5.40 Log.结论 建立的rhGH-Fc融合蛋白灭活/去除工艺可有效灭活/去除未知的、新出现的病毒.
Verification of virus inactivation/removal efficacy in pilot-scale procedure of recombinant human growth hormone-Fc
Objective To verify the virus inactivation/removal efficacy in pilot-scale procedure of recombinant human growth hormone-Fc(rhGH-Fc)fusion protein in order to ensure the product quality and patient safety.Methods Four viruses,pseu-dorabies virus(PRV),xenotropic murine leukemia virus(X-MuLV),minute virus of mice(MVM),and reovirus-3(Reo-3),were used as indicator viruses.Under the condition of scaling down,the inactivation ability of solvent and detergent(S/D)method on PRV and X-MuLV,the removal ability of X-MuLV and MVM by affinity chromatography,and the removal ability of MVM and Reo-3 by nanofilm filtration,were investigated.Results The inactivation ability of PRV and X-MuLV by S/D method was ≥ 4.95 and＞5.15 Log,respectively.The removal ability of X-MuLV and MVM by affinity chromatography was ≥ 4.36 and 1.68 Log,respectively.The removal capacity of MVM and Reo-3 by nanofilm filtration was＞5.36 and＞5.40 Log,respectively.Conclusion The established rhGH-Fc fusion protein inactivation/removal process can effectively inactivate/remove the unknown and emerging viruses.
Recombinant human growth hormone-Fc(rhGH-Fc)Virus inactivationVirus removalSolvent and detergent(S/D)methodAffinity chromatographyNanofilm filtration
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