| Popis: |
Adenovirus belongs to the family of Adenoviruses, as a vaccine carrier, it not only has high safety, but also can stimulate the body to produce cellular immunity and humoral immunity. In this study, We aimed to prepare the adenoviral vectors specific single domain antibody to use in adenovirus identification and purification. We successfully constructed a single domain antibody phage display library with the capacity of 1.8×109 by immunizing and cloning the VHH gene from the Bactrian camel. After second round of biopanning, the clones specific for adenovirus were screened by Phage ELISA. Twenty-two positive clones were obtained, and two clones with the highest binding affinity from ELISA were selected and named as sdAb 5 and sdAb 31 respectively for further application. The recombinant single domain antibody was soluble expressed in E. coli, and can specifically bind to adenovirus rAd26, ChAd63 and Had5 in ELISA assay and in live cell immunofluorescence. At last, we established an effective method for immunoaffinity purification of adenovirus by immobilizing the single domain antibody to Sepharose beads, and it can be used to selectively capture Adenoviruses from cell culture medium. The preparation of the adenovirus specific single domain antibody was laying a foundation for the one-step immunoaffinity purification and identification of Adenoviruses. |
