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目的采用层析技术制备高纯度的马抗中东呼吸综合征冠状病毒(Middle East respiratory syndrom coronavirus,MERS-CoV)F(ab’)_2。方法以MERS-CoV病毒样颗粒(virus-like particles,VLPs)对马匹进行多次免疫所制备的高免血清为原料,采用亲和层析技术提取血清IgG,经胃蛋白酶酶切后,通过阴离子交换层析、疏水层析、凝胶过滤层析分别对F(ab’)_2进行纯化;以MERS假病毒检测所制备的IgG与F(ab’)_2中和活性。结果凝胶过滤层析实现了F(ab’)_2的高纯度分离,最终制备的F(ab’)_2收率约70%,纯度为93%以上,具有良好的中和活性,其半数抑制浓度(IC_(50))为5.13μg/ml。结论制备了高纯度马抗MERS-CoV免疫球蛋白F(ab’)_2,为人用抗MERS-CoV免疫球蛋白F(ab’)_2生产工艺的建立奠定了基础。
OBJECTIVE To prepare high-purity MERS-CoV F (ab ’) _ 2 by chromatography. Methods Serum IgG was extracted from the serum of MERS-CoV virus-like particles (VLPs) by multiple immunization. The serum IgG was extracted by pepsin, The purified F (ab ’) _ 2 was purified by chromatography, hydrophobic chromatography and gel filtration chromatography. The neutralizing activity of IgG and F (ab’) _ 2 was tested by MERS pseudovirus. Results F (ab ’) _ 2 was purified by gel filtration chromatography. The final yield of F (ab’) _ 2 was about 70% with a purity of over 93%. It had good neutralization activity and half inhibition The concentration (IC 50) was 5.13 μg / ml. Conclusion High purity horse anti-MERS-CoV immunoglobulin F (ab ’) _ 2 was prepared, which laid the foundation for the establishment of human anti-MERS-CoV immunoglobulin F (ab’) _ 2 production technology.