Valdés R, Ibarra Neysi, Ruibal I, Beldarraín A, Noa E, Herrera N, Alemán R, Padilla S, Garcia J, Pérez M, Morales R, Chong E, Reyes B, Quiñones Y, Agraz A, Herrera L
Center for Genetic Engineering and Biotechnology, Ave 31/158 and 190, PO Box 6162, Havana 10600, Cuba.
J Biotechnol. 2002 Jul 3;96(3):251-8. doi: 10.1016/s0168-1656(02)00047-0.
The virus removal of protein A affinity chromatography, inactivation capacity, acid pH and a combination of high temperature with a chaotropic agent was determined in this work. The model viruses studied were sendaivirus, human immunodeficency virus (HIV-IIIb), human poliovirus type-II, human herpesvirus I and canine parvovirus. The protein A affinity chromatography showed a maximum reduction factor of 8 logs in the case of viruses larger than 120 nm size, while for small viruses (18-30 nm) the maximum reduction factor was about 5 logs. Non viral inactivation was observed during the monoclonal antibody elution step. Low pH treatment showed a maximum inactivation factor of 7.1 logs for enveloped viruses. However, a weak inactivation factor (3.4 logs) was obtained for DNA nonenveloped viruses. The combination of high temperature with 3 M KSCN showed a high inactivation factor for all of the viruses studied. The total clearance factor was 23.1, 15.1, 13.6, 20.0 and 16.0 logs for sendaivirus, HIV-IIIb, human poliovirus type-II, human herpesvirus I and canine parvovirus, respectively.
本研究测定了蛋白A亲和层析的病毒去除效果、灭活能力、酸性pH以及高温与离液剂联合使用的效果。所研究的模型病毒有仙台病毒、人类免疫缺陷病毒（HIV-IIIb）、人类Ⅱ型脊髓灰质炎病毒、人类疱疹病毒I和犬细小病毒。对于尺寸大于120nm的病毒，蛋白A亲和层析显示出最大8个对数的降低因子，而对于小病毒（18-30nm），最大降低因子约为5个对数。在单克隆抗体洗脱步骤中观察到非病毒灭活现象。低pH处理对包膜病毒显示出最大7.1个对数的灭活因子。然而，对于DNA非包膜病毒，获得的灭活因子较弱（3.4个对数）。高温与3M KSCN联合使用对所有研究的病毒都显示出高灭活因子。仙台病毒、HIV-IIIb、人类Ⅱ型脊髓灰质炎病毒、人类疱疹病毒I和犬细小病毒的总清除因子分别为23.1、1 '>本研究测定了蛋白A亲和层析的病毒去除效果、灭活能力、酸性pH以及高温与离液剂联合使用的效果。所研究的模型病毒有仙台病毒、人类免疫缺陷病毒（HIV-IIIb）、人类Ⅱ型脊髓灰质炎病毒、人类疱疹病毒I和犬细小病毒。对于尺寸大于120nm的病毒，蛋白A亲和层析显示出最大8个对数的降低因子，而对于小病毒（18-30nm），最大降低因子约为5个对数。在单克隆抗体洗脱步骤中观察到非病毒灭活现象。低pH处理对包膜病毒显示出最大7.1个对数的灭活因子。然而，对于DNA非包膜病毒，获得的灭活因子较弱（3.4个对数）。高温与3M KSCN联合使用对所有研究的病毒都显示出高灭活因子。仙台病毒、HIV-IIIb、人类Ⅱ型脊髓灰质炎病毒、人类疱疹病毒I和犬细小病毒的总清除因子分别为23.1、15.1、13.6、20.0和16.0个对数。 5、13.6、20.0和16.0个对数。
