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目的制备肝水解肽,并对其病毒灭活/去除工艺进行验证。方法以鸡新城疫病毒(NDV)和鸡传染性法氏囊病毒(IBDV)为指示病毒,在100℃,pH6.0～7.0煮沸20min灭活病毒,用截留相对分子质量为10000的中空纤维柱,在进压0.15Mpa、回流压0.05Mpa和进压0.20Mpa、回流压0.10Mpa各循环超滤3次去除病毒,用无特定病原(SPF)的鸡胚和鸡成纤维细胞进行病毒灭活/去除的效果验证。结果加热煮沸后,指示病毒未检出,滴度降低NDV≥7.4logEID50/0.2ml、IBDV≥5.45logCCID50/0.1ml;超滤后IBDV未检出,滴度降低≥5.43logCCID50/0.1ml。盲传复壮均未检出病毒。制备的肝水解肽多肽含量均在20mg/ml以上。结论制备的肝水解肽工艺中两种灭活/去除病毒方法均有效,经灭活/去除病毒后的产品质量稳定。
Objective To prepare hepatic hydrolyzate peptide and verify its virus inactivation / removal process. Methods NDV and IBDV were used as the indicator virus to inactivate the virus by boiling at 100 ℃, pH 6.0-7.0 for 20 minutes. The recombinant plasmids were stained with hollow fiber column with relative molecular mass of 10,000. Pressure 0.15Mpa, reflux pressure 0.05Mpa and pressure 0.20Mpa, back pressure 0.10Mpa each cycle of ultrafiltration three times to remove the virus, with no specific pathogen (SPF) of chicken embryo and chicken fibroblasts for virus inactivation / removal Effect verification. Results After boiling, the virus was not detected. The titer of NDV≥7.4logEID50 / 0.2ml and IBDV≥5.45logCCID50 / 0.1ml were detected. After the ultrafiltration, the IBDV was not detected, and the titer was decreased by5.43logCCID50 / 0.1ml. Blind rejuvenation have not detected the virus. Preparation of liver hydrolyzed peptide content of more than 20mg / ml. Conclusion The two methods of inactivation / removal of virus in the preparation of hepatic hydrolytic peptide are effective, and the quality of the product after inactivation / removal of virus is stable.