马抗SARS-CoV-2特异性IgG与F(ab’)2的制备研究  

Preparation of equine anti-SARS-CoV-2 immunoglobulin special IgG and special F(ab’)_(2)

作　　者：许梦园 毕津豪 王馨悦 张颖[2,5] 韩秋雪[2,6] 莫若 闫飞虎 杨松涛 冯娜 王铁成[2] 赵永坤 夏咸柱[2,1,3] XU Meng-yuan;BI Jin-hao;WANG Xin-yue;ZHANG Ying;HAN Qiu-xue;MO Ruo;YAN Fei-hu;YANG Song-tao;FENG Na;WANG Tie-cheng;ZHAO Yong-kun;XIA Xian-zhu(College of Veterinary Medicine,Jilin University,Changchun 130062,China;Changchun Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Changchun 130112,China;College of Veterinary Medicine,Jilin Agricultural University,Changchun 130118,China;College of Veterinary Medicine,Northeast Agricultural University,Harbin 150030,China;College of Wildlife and Protected Area,Northeast Forestry University,Harbin 150040,China;Beijing Union Medical College Comparative Medical Center,Beijing 100021,China)

机构地区：[1]吉林大学动物医学学院,吉林长春130062 [2]中国农业科学院长春兽医研究所,吉林长春130112 [3]吉林农业大学动物医学院,吉林长春130118 [4]东北农业大学动物医学学院,黑龙江哈尔滨150030 [5]东北林业大学野生动物与自然保护地学院,黑龙江哈尔滨150040 [6]北京协和医学院比较医学中心,北京100021

出　　处：《中国兽医科学》2022年第7期846-852,共7页Chinese Veterinary Science

摘　　要：收集经SARS-CoV-2病毒样颗粒(VLPs)为免疫原免疫的健康马血清,采用亲和层析法分离血清中总IgG,利用胃蛋白酶消化Fc片段获得免疫球蛋白F(ab’)_(2),将二者使用结合RBD蛋白的免疫层析柱进行纯化,获得具有高度特异性的马抗SARS-CoV-2 IgG与免疫球蛋白F(ab’)_(2)。结果显示,特异性IgG与F(ab’)_(2)浓度是2.36 mg/mL和1.05 mg/mL,收率为11%和4.89%,纯度在91%与96%以上,对假病毒的半抑制浓度(IC50)为1.406μg/mL和0.862μg/mL。结果表明,成功制备了纯度高、特异性强且具有中和活性的马抗SARS-CoV-2特异性IgG与特异性F(ab’)_(2),为制备安全高效的抗SARS-CoV-2治疗性抗体药物奠定了基础。After immunizing healthy horses with SARS-CoV-2 virus-like particles(VLPs)as immunogens,immunized horse serum was collected.The total IgG in the serum was separated by affinity chromatography,and then digested with pepsin to obtain immunoglobulin F(ab’)_(2),the IgG and F(ab’)_(2) using an immunochromatographic column that binds to the RBD protein to obtain a highly specific horse Anti-SARS-CoV-2 IgG and F(ab’)_(2).It's concentration of IgG and F(ab’)_(2) is 2.36 mg/mL and 1.05 mg/mL,while the recovery rates were 11%and 4.89%,and the purities of prepared IgG and F(ab’)_(2) were 91%and 96%.Semi-inhibited concentrations of pseudovirus(IC50)were 1.406μg/mL and 0.862μg/mL.These results show that a high purity,specificity,activity of specific IgG and F(ab’)_(2) against SARS-CoV-2 was prepared successfully,which laid a foundation for preparing safe and efficient anti-SARS-CoV-2 therapeutic antibody drugs.

关 键 词：新型冠状病毒 马免疫球蛋白 F(ab’)2 亲和层析 假病毒中和试验 

分 类 号：S852.659.6[农业科学—基础兽医学]