抗甲肝病毒人源基因工程全抗体分子在杆状病毒中的表达  

Baculovirus expression of two human recombinant neutralizing IgG monoclonal antibodies to hepatitis A virus

作　　者：曹经瑗[1] 梁米芳[2] 孟庆玲[1] 王晓芳[2] 许永刚[2] 郭可謇[1] 詹美云[1] 毕胜利[1] 李德新[2] 

机构地区：[1]中国疾病控制中心病毒病预防控制所肝炎室 [2]中国疾病控制中心病毒病预防控制所出血热脑炎室

出　　处：《中华实验和临床病毒学杂志》2004年第1期20-23,共4页Chinese Journal of Experimental and Clinical Virology

摘　　要：目的　探讨人源抗甲型肝炎病毒全抗体分子在杆状病毒中的表达。方法　将获得的人源抗甲肝病毒中和性抗体Fab段基因克隆入含信号肽及Fc的杆状病毒表达载体中并在杆状病毒细胞中表达。结果　获得了中和性人源抗甲肝病毒全抗体分子的表达产物并进行了纯化 ,轻重链表达产物位置大小正确 ,HAFc16抗体能与具有中和活性的鼠抗甲肝病毒单克隆抗体产生竞争抑制反应 ,并能在体外中和甲肝病毒 ,另一株HAFc78抗体同样具有体外中和甲肝病毒的活性 ,但系抗不同位点的抗体。结论　获得的人源抗甲肝病毒全抗体分子表达产物具有很好的体外中和甲肝病毒的活性 ,且为抗不同位点的抗体 ,为这些抗体的进一步开发及应用打下了基础 ,为防止甲型肝炎暴发流行提供应急措施。Objective To develop human recombinant neutralizing IgG monoclonal antibodies to hepatitis A virus (HAV) by baculovirus expression system. Methods The heavy and light chain genes of two human-derived neutralizing Fab antibodies to HAV were cloned into baculovirus expression vector PAc-κ-Fc and PAc-L-Fc, and further expressed in insect cells as IgG antibodies. The IgG products were purified and well characterized. Results The baculovirus expressed McAb HAFc16 fully retained the specificity of binding to hepatitis A virus and the competition with mouse anti-hepatitis A virus McAb using ELISA. The viral neutralization assay in vitro demonstrated the retention of antibody function after expression of the human antibody in insect cells. The other expressed antibody HAFc78 also has the neutralizing activity but it is directed against different epitopes of HAV when compared with HAFc16. Conclusion The recombinant baculovirus/insect cells expressed human neutralizing IgG antibodies to hepatitis A virus retained all biological functions specific for hepatitis A virus. The results provided the possibility of using these antibodies to rapidly protect high risk or early exposure populations from hepatitis A virus infection.

关 键 词：甲肝病毒 基因工程 杆状病毒 肝炎抗体 全抗体分子 

分 类 号：R392[医药卫生—免疫学] R373[医药卫生—基础医学]