人類微小病毒B19-VP1u單株抗體製備與應用

人類微小病毒B19 (Human Parvovirus B19, B19)會造成許多疾病，包括貧血、心肌炎和自體免疫疾病等。至目前為止，對於B19病毒引起的疾病並沒有治療藥物和疫苗可以施打。許多文獻已證明B19病毒殼體蛋白獨立區域(B19-VP1u)在B19病毒成功感染宿主細胞時扮演關鍵角色。因此本論文之目的在製備單株抗體可以辨認B19-VP1u，篩選可以生產辨認B19-VP1u病毒抗體之單株細胞株，更進一步對單株抗體的種類及辨認之抗原性分析，然後再測定這些單株抗體對其他B19病毒蛋白之交叉反應性。實驗結果獲得兩株可作用於B19-VP1u病毒之單株抗體，分別命名為3GC11F3及3GC11F9。3GC11F3單株抗體的效價可達5萬至10萬倍，3GC11F9單株抗體的效價可達1萬至5萬倍。單株抗體抗原結合利用競爭型酵素免疫分析法評估抗體專一性。確認3GC11F3單株抗體分型IgG1 kappa light chain及3GC11F9單株抗體分型IgG1 kappa light chain和IgG3 kappa light chain。

Human Parvovirus B19 (B19) caused a variety of diseases, including anemia, myocarditis and autoimmune disease. So far, there is no treatment for the disease caused by B19 and no vaccine to prevent the infection. In many researches has found that B19 virus infected cell and the B19-VP1u protein played an important role in B19 infection. Thus, in this study, we produce B19-VP1u specific monoclonal antibody, characterize the properties of the antibody and test the antibody if cross reaction to other B19 virus protein. We got two B19-VP1u hybridoma clones, named 3GC11F3 and 3GC11F9. The titer of 3GC11F9 is between 10000 to 50000x and 3GC11F3 is between 50000 to 100000x. Antibody binding was evaulated using competitive ELISA assay. The isotype of 3GC11F3 monoclonal antibody is IgG1 kappa light chain. The isotype of 3GC11F9 monoclonal antibody is IgG1 kappa light chain and IgG3 kappa light chain.