The whole DNA-Launched infectious GSK690693 of DHAV-1 named pIR-DHAV-1 was completed by respectively inserting four fragments of strain LY0801 into pIR vector with multistep strategy. The complete VP1 gene of DHAV-2 04G strain (EF067923.1) and all of the primers were synthesized by Sangon Biotech Co., Ltd. (Shanghai, China).
2.2. Neutralization assay for mAb 4F8 against DHAV-1 and DHAV-3
In this study, mAb 4F8 against DHAV-1 was quantified as 2.056 mg/mL. With 2, 4, 8, 16, 32 and 64 dilution in saline, the neutralizing activities of mAb 4F8 against DHAV-1 and DHAV-3 were determined using quantitative virus (100 ELD50, 0.1 mL) respectively.
2.3. Preliminary analysis of linear epitope to mAb 4F8
Table 1.
Primers for construction of recombinant bacmids.Protein namePrimersSequence (5′–3′)Size (bp)1BP11FCGCGGATCCATGGATACTCTTACCAAAAAC21931RAACCTCGAGTCATTCAATTTCCAAATTGAGTTCG1BVP01FCGCGGATCCATGGATACTCTTACCAAAAAC7681R0CGGCTCGAGTCATCTGATTGTCAAATGGTC1BVP31F3CCGGGATCCATGGGAAAGAGAAAACCACG7111R3CCGCTCGAGTCACTGATTATTGGTTGCCATCTGC1BVP11F1ATTGGATTCATGGGTGATTCTAACCAGTTAGGGGATG7141RAACCTCGAGTCATTCAATTTCCAAATTGAGTTCG2BVP12F1AGTGAATTCACCATGGGGGACACTAACCAGC7132R1GCGCTCGAGCTATTCAATTTCAAGATGGAGC3BP13FATTGGATCCATGGGAGCTGTTGAGAGTG21093RACGCTCGAGTCATTCAATTTCCAAA3BVP03FATTGGATCCATGGGAGCTGTTGAGAGTG6783R0ATACTCGAGTCACTGGTCATTGAAGGGC3BVP33F3ATAGGATTCATGGGTAAGAAGAAACC7113R3ATACTCGAGTCACTGGTTATTGGCG3BVP13F1ACTGGATTCATGGGTGATTCTAATC7203RACGCTCGAGTCATTCAATTTCCAAAFull-size tableTable optionsView in workspaceDownload as muscular system CSV
2.2. Neutralization assay for mAb 4F8 against DHAV-1 and DHAV-3
In this study, mAb 4F8 against DHAV-1 was quantified as 2.056 mg/mL. With 2, 4, 8, 16, 32 and 64 dilution in saline, the neutralizing activities of mAb 4F8 against DHAV-1 and DHAV-3 were determined using quantitative virus (100 ELD50, 0.1 mL) respectively.
2.3. Preliminary analysis of linear epitope to mAb 4F8
Table 1.
Primers for construction of recombinant bacmids.Protein namePrimersSequence (5′–3′)Size (bp)1BP11FCGCGGATCCATGGATACTCTTACCAAAAAC21931RAACCTCGAGTCATTCAATTTCCAAATTGAGTTCG1BVP01FCGCGGATCCATGGATACTCTTACCAAAAAC7681R0CGGCTCGAGTCATCTGATTGTCAAATGGTC1BVP31F3CCGGGATCCATGGGAAAGAGAAAACCACG7111R3CCGCTCGAGTCACTGATTATTGGTTGCCATCTGC1BVP11F1ATTGGATTCATGGGTGATTCTAACCAGTTAGGGGATG7141RAACCTCGAGTCATTCAATTTCCAAATTGAGTTCG2BVP12F1AGTGAATTCACCATGGGGGACACTAACCAGC7132R1GCGCTCGAGCTATTCAATTTCAAGATGGAGC3BP13FATTGGATCCATGGGAGCTGTTGAGAGTG21093RACGCTCGAGTCATTCAATTTCCAAA3BVP03FATTGGATCCATGGGAGCTGTTGAGAGTG6783R0ATACTCGAGTCACTGGTCATTGAAGGGC3BVP33F3ATAGGATTCATGGGTAAGAAGAAACC7113R3ATACTCGAGTCACTGGTTATTGGCG3BVP13F1ACTGGATTCATGGGTGATTCTAATC7203RACGCTCGAGTCATTCAATTTCCAAAFull-size tableTable optionsView in workspaceDownload as muscular system CSV