钴^(60)-γ射线辐照法消毒冻干免疫球蛋白的效果观察

OBSERVATION ON EFFICACY OF ^(60)Co-γ IRRADIATION IN DISINFECTION OF LYOPHILIZED IMMUNOGLOBULIN

目的探讨钴60-γ射线辐照消毒法处理冻干免疫球蛋白的可行性。方法采用病毒定量检测方法,观察钴60-γ射线辐照法对免疫球蛋白中病毒灭活效果。结果含病毒的免疫球蛋白经30kGy剂量钴60辐照处理,对含不同蛋白保护剂的冻干IgG制品中辛德比斯病毒灭活对数值≥5.5TCID50。经30kGy剂量辐照灭菌处理,冻干IgG制品外观未见明显改变,各处理组之间的pH值与未处理组结果相近。蛋白电泳Native-PAGE分析各保护剂组之间无明显差别,保护剂浓度高低对试验结果无明显影响;SDS-PAGE分析,各处理组中的IgG经还原后其重链和轻链仍清晰可见,各保护剂组结果间无明显差别。高效液相色谱法检测结果显示,三种保护剂对IgG分子的保护效果较好,分子聚合和断裂峰明显减少。冻干IgG经辐照处理后抗原含量与对照相比均达80%以上,各保护剂组之间抗原含量基本一致。结论冻干血浆蛋白制品IgG经γ-射线照射后能够保持一定的结构和功能的完整性。

Objective To explore the feasibility of disinfection of lyophilized immunoglobulin with ^6Co-γ irradiation. Methods Virus quantitative detection method was used to observe the efficacy of ^6Co-γ irradiation in inactivating virus in immunoglobulin. Results Treatment of the immunoglobulin containing virus with 30 kGy of ^6Co-γ irradiation inactivated Sindbis virus in the lyophilized IgG preparation containing different protein protectives by a logarithm value t〉 5.5 TCID50. After treatment with 30 kGy of radiation sterilization, the external appearance of the lyophilized IgG preparation did not change significantly, the pH values between different treated groups and between treated and non -treated groups were approximate. Native - PAGE analysis of protein electrophoresis analysis results between the groups containing different pro- tectives had no significant difference and the concentrations of protectives had no significant influence on the test results. SDS - PAGE analysis showed that the heavy chain and light chain of IgG in different treated groups still could be seen clearly after reduction and there was no significant difference between the results of the groups containing different protectives. High performance liquid chromatography results showed that the efficacy of three protectives in protecting IgG molecules was good and polymerization and break peak of the molecules reduced significantly. The antigen content of the lyophilized IgG reached over 80% after radiation treatment as compared with the control group and the antigen contents of the groups containing different protectives were essentially identical. Conclusion The lyophilized plasma protein preparation IgG can maintain certain structural and functional integrity after γ -irradiation.